Vaginal isolation of beta‐haemolytic Streptococcus from bitches with and without neonatal deaths in the litters

The aim of the study was to identify beta‐haemolytic streptococci in the vagina of bitches who had delivered healthy litters and bitches who had delivered litters in which neonatal deaths occurred. Fifty‐one bitches divided into two groups were used. Group 1 (G1) included 28 bitches that had delivered healthy litters and group 2 (G2) included 23 bitches that had delivered puppies who died in the neonatal period. Two vaginal samples were taken, one in proestrus and the other at the end of gestation (EG). Beta‐haemolytic Streptococcus (BS) was isolated from 16 bitches (57%) in G1 and from 21 bitches (91%) in G2. The bacteriological cultures, serological tests (Streptex^®) and PCR assay allowed identification of Streptococcus canis and Streptococcus dysgalactiae in G1 and G2. Ultramicroscopic studies allowed the observation of M Protein and capsules in strains of S. dysgalactiae and S. canis in G1 and G2. The S. canis strains isolated from G2 showed thicker capsules than S. canis strains isolated from G1 (234 ± 24.2 vs 151.23 ± 28.93 nm; p < .001.). No differences were observed in capsule thickness between strains of S. dysgalactiae isolated from G1 and G2 (210 ± 13.54 vs 211.66 ± 19.67 nm; p > .70). All strains of beta‐haemolytic Streptococcus isolated were penicillin sensitive. Penicillin was administered from EG to 5 days post‐partum in 10 G2 females with isolation of BS (G2A). Saline solution was administered in eleven G2 females with isolation of BS (G2B). Ninety per cent of the puppies survived in G2A and 25% survived in G2B. Our results suggest BS is involved in canine neonatal deaths.     

Retracted: Effect of Post‐Thaw Storage Time on Motility and Fertility of Cryopreserved Beluga Sturgeon (Huso huso) Sperm

The aim of this study was to test the influence of post ‐ thaw storage time on the duration of sperm motility, percentage of motile sperm, and fertilization and hatching rates of fresh sperm and sperm stored for 0, 30 and 60 min at 4°C post‐thawing. After being frozen in liquid nitrogen and then thawed, the percentage of motile sperm and duration of motility were not affected by 30 min of storage at 4°C, whereas a significant decline in these parameters was observed after 60 min of storage. Similarly, fertilization and hatching rates were significantly affected within 60 min of storage at 4°C, and the fertility of frozen‐thawed sperm was significantly lower than that of fresh sperm. We conclude that cryopreserved sperm of beluga sturgeon could be stored for 30 min without the loss of sperm quality. This described procedure for beluga sturgeon cryopreservation is reliable and efficient and therefore can be recommended for hatchery practice after scaling up this technique.     

Neonatal Clinical Evaluation, Blood Gas and Radiographic Assessment After Normal Birth, Vaginal Dystocia or Caesarean Section in Dogs

This study aimed to standardize signs and diagnostic criteria of respiratory function in newborn puppies delivered normally or after dystocia and caesarean operation. A total of 48 neonates were allocated into groups: eutocia (n = 20), dystocia (n = 8), caesarean (c)-section (n = 20). Neonatal health was assessed using the Apgar score and body temperature was determined at 0, 5 and 60 min after delivery. Venous blood gases (pO₂ and SO₂) was measured immediately and 60 min after delivery, and a thoracic radiograph was made between 0 and 5 min of life. The c-section group had significantly lower Apgar scores at birth and 5 min. Hypothermia was present at 5 min in the eutocia and c-section groups, and at 60 min in all groups. The eutocia group had an irregular respiratory pattern in 78% of puppies at birth, 27.7% at 5 min and 21% at 60 min compared with 87.5%, 62.5% and 12.5% of the pups in the dystocia group where there was irregular respiratory rhythm, moderate to intense respiratory sounds with agonic episodes. The c-section group had respiratory alterations in 70%, 45% and 16% of puppies at 0, 5 and 60 min, respectively. Radiographic abnormalities were present in 17% of the pups in the eutocia group, 25% of the pups in the dystocia group and 30% of the pups in the c-section group, respectively. The c-section group had significantly lower SO₂ values at 60 min than at birth. All puppies had hypoxaemia, but a significant decrease was observed in the c-section group. Newborn puppies had tissue hypoxia and irregular respiratory pattern at birth. Caesarean-section puppies had lower vitality; however, all developed satisfactory Apgar scores at 5 min of life, regardless of the obstetric condition.     

Arachidic Acid in Extender Improves Post‐thaw Parameters of Cryopreserved Nili‐Ravi Buffalo Bull Semen

Cryopreservation process reduces lipids and phospholipids from buffalo bull spermatozoa. It was therefore hypothesized that supplementation of fatty acid to extender may improve the post‐thaw quality of buffalo semen. The objective was to evaluate the effect of arachidic acid supplementation in extender on post‐thaw quality of buffalo bull (Bubalus bubalis) spermatozoa. Semen was collected from three adult Nili‐Ravi buffalo bulls of similar age group with artificial vagina (42°C) for 3 weeks (replicate). Qualified semen ejaculates (n = 18) were split into four aliquots and diluted in tris–citric acid extender containing 0.0 (control), 5.0, 10.0 and 20.0 ng/ml at 37°C having approximately 50 × 10⁶ spermatozoa/ml. Diluted semen was cooled to 4°C in 2 h and equilibrated for 4 h at 4°C. Cooled semen was filled in 0.5‐ml straws at 4°C, kept on liquid nitrogen vapours for 10 min and plunged in liquid nitrogen for storage. Thawing of frozen semen was performed after 24 h at 37°C for 30 s. Sperm progressive motility (%) was improved in a dose‐dependent manner by supplementing arachidic acid at 5.0, 10.0 and 20.0 ng/ml compared with control. Structural and functional integrity of sperm plasma membrane (%), number of acrosome‐intact live sperm (%) and sperm chromatin integrity (%) were better (p < 0.05) in extender having 5.0 ng/ml of arachidic acid compared with control. At 10.0 ng/ml, these values did not vary (p > 0.05) from those at 5.0 ng/ml. Further improvement in structural and functional integrity of sperm plasma membrane, number of acrosome‐intact live sperm and chromatin integrity was observed at 20.0 ng/ml of arachidic acid in extender. In conclusion, arachidic acid supplementation in extender improved the post‐thaw quality parameters of cryopreserved Nili‐Ravi buffalo bull spermatozoa. Among the arachidic acid concentrations studied, maximum improvement in post‐thaw semen quality parameters was observed at 20.0 ng/ml.     

The Oxidative Stress,Antioxidant Profile and Acid–base Status in Preterm and Term Canine Neonates

During the initiation of neonatal pulmonary respiration, there is an exponential increase in reactive oxygen species that must be scavenged by antioxidant defences. However, neonate and preterm newborns are known to possess immature antioxidant mechanisms to neutralize these toxic effects. The purposes of this study were to compare the development of antioxidant system between preterm and term canine neonates and to evaluate the magnitude of acid–base balance during the initial 4 h of life. A prospective study was conducted involving 18 neonatal puppies assigned to Term Group (63 days of gestation; n = 5), Preterm‐57 Group (57 days of gestation; n = 8) and Preterm‐55 Group (55 days of gestation; n = 5). Neonates were physically examined through Apgar score and venous haemogasometry within 5 min, 2 and 4 h after birth. No difference on amniotic fluid and serum superoxide dismutase (SOD), glutathione peroxidase (GPx) and the marker of oxidative stress (thiobarbituric acid reactive substances; TBARS) was verified. Irrespective of prematurity, all neonates presented low vitality, hypothermia, acidosis, hypoxaemia and hypercapnia at birth. However, term puppies clinically evolved more rapidly than preterm newborns. During the course of the study, premature neonates presented more severe complications, such as prolonged hypoxaemia and even death. In conclusion, premature puppies have no signs of immature enzymatic mechanisms for controlling oxidative stress, although SOD and GPx may participate in achieving acid–base balance. Aside from initial unremarkable symptoms, premature puppies should be carefully followed up, as they are at high risk of succumbing to odds of prematurity.     

Fasting heat production of Saanen and Anglo Nubian goats measured using open‐circuit facemask respirometry

This study aimed to establish the heat production (HP) of Saanen and Anglo Nubian goats at absorptive (feeding) and at post‐absorptive (fasting) statuses to determine the adequate period of fasting required for the measurement of basal metabolism. Gas exchange was recorded via open‐circuit facemask respirometry. Six non‐lactating and non‐pregnant goats of each breed, Saanen (49.2 ± 3.2 kg of body weight, BW) and Anglo Nubian (64.0 ± 3.0 kg BW), were placed in individual pens with ad libitum access to the same total mixed ration. After a 3‐day feeding period, the animals were subjected to fasting (no feed), and the gas exchange measurement was performed for 30 min at 0, 12, 20, 36, 44, 60 and 68 h after fasting. The daily HP of the Saanen and Anglo Nubian goats averaged 557.4 ± 38.7 and 357.1 ± 35.3 kJ/kg^0.75 BW day respectively. During fasting, the methane production decreased exponentially in both breeds, and the critical time when methane production was statistically equal to zero was at 31 h of fasting for the Saanen goats and at 40 h for the Anglo Nubian goats. The daily HP and respiratory exchange rate during fasting decreased up to 60 h. Taken together, our results suggest that the ideal period to measure fasting heat production (FHP) for goats fed at maintenance levels should be between 40 h and 60 h of fasting. Consequently, the daily FHP, after 60 h of fasting, of Saanen and Anglo Nubian goats was 183.3 ± 16.3 and 211.1 ± 11.5 kJ/kg^0.75 BW day respectively. The results presented herein are relevant for future studies of energy metabolism in goats.     

Effects of L‐citrulline supplementation on heat stress physiology,lactation performance and subsequent reproductive performance of sows in summer

Lactating sows are susceptible to heat stress (HS). Part of the thermoregulatory response to HS is to increase peripheral blood flow, which is mediated in part by the vasodilator, nitric oxide (NO). Therefore, the aim of this experiment was to determine the effect of supplementation of L‐citrulline, a NO precursor, on symptoms of HS, lactation performance and subsequent reproductive performance of sows in summer. A total of 221 summer farrowing mixed parity sows were fed either a control diet or supplemented with 1% L‐citrulline upon entry to the farrowing house (6 ± 1.8 days for mean ± standard deviation [SD] before farrowing) until weaning (26 ± 1.5 days). The average daily minimum and maximum temperature in the farrowing house was 21.0 ± 1.88 and 29.2 ± 3.82°C (mean ± SD). Rectal temperature, respiration rate, and plasma and urinary nitrite and nitrate (NOx) of sows were measured on the 19th day post‐farrowing. Supplemental L‐citrulline in the diet did not affect the number of piglets born alive, feed intake of sows, body weight or backfat thickness of sows at weaning, or litter weight gain. L‐citrulline tended to reduce piglet pre‐weaning mortality rate from 18.6% to 15.6% (p = 0.058). L‐citrulline reduced the respiration rate of sows compared to the control diet at 17:00 hr (Time × Diet, p < 0.001); however, rectal temperature was not affected. L‐citrulline tended to increase urinary NOx concentrations (127 vs. 224 µM, p = 0.057) but not plasma NOx concentrations. L‐citrulline did not affect farrowing rate or number of piglets born alive in the subsequent parity. In conclusion, L‐citrulline supplementation reduced respiration rate of lactating sows and reduced piglet pre‐weaning mortality rate in summer. Whether the effects were due to a NO‐dependent mechanism requires further validation.     

Blood Metabolite Profiles in Cycling and Non‐cycling Friesian–Sanga Cross‐bred Cows Grazing Natural Pasture During the Post‐partum Period

An experiment was conducted to investigate the effect of plasma concentrations of the metabolic hormones [Growth hormone (GH), insulin and insulin‐like growth factor –I (IGF‐I)] and nutritional metabolites (Glucose, cholesterol, total protein, albumin, globulin, urea and creatinine) on the resumption of post‐partum ovarian activity in sixteen Friesian–Sanga cows grazing extensively on native grassland. Blood samples were taken from cows from week 1 to 16 post‐partum. Cows were classified as having resumed ovarian activity when a plasma progesterone concentration of ≥ 1.0 ng/ml was recorded for two consecutive weekly samples. Based on the resumption of ovarian activity, cows were classified as early‐cycling, late‐cycling or non‐cycling. The concentrations of the metabolic hormones were measured from week 1 to 10, while those of the nutritional metabolites were measured during week 1, 3, 5, 7 and 9 during the study period. The concentrations of the metabolic hormones, GH and insulin were similar (p > 0.05) in the three ovarian activity groups, likewise the concentrations of the nutritional metabolites, glucose, total protein, globulin, urea and creatinine. Plasma IGF‐I concentration was higher (p < 0.001) in early‐cycling (18.7 ± 0.74 ng/ml) than in late‐cycling (12.4 ± 0.75 ng/ml) and non‐cycling (10.4 ± 0.91 ng/ml) cows. Plasma cholesterol concentrations were significantly lower (p < 0.05) in early‐cycling (1.94 ± 0.15 mmol/l) compared with late‐cycling (2.48 ± 0.12 mmol/l) and non‐cycling (2.61 ± 0.11 mmol/l) cows. For plasma albumin concentrations, the levels recorded for early‐cycling cows were higher (40.7 ± 2.85 g/l) than in late‐cycling (34.4 ± 1.97 g/l) and non‐cycling (33.6 ± 2.66) cows. The results suggest that cows with lower plasma concentrations of IGF‐I and albumin, but higher plasma cholesterol concentrations were at risk of delayed resumption of post‐partum ovarian activity.     

Orally administered Chrysin improves post‐thawed sperm quality and fertility of rooster

Chrysin is a bioflavonoid compound found in passion flower, chamomile, propolis and honey at high levels. Post‐thawed sperm quality and fertility of Chrysin‐fed roosters were assessed in this study. Twenty 40‐week‐old male broiler breeders were randomly divided into four groups and fed basal diet supplemented with different levels of Chrysin including 0 (Ch‐0), 25 (Ch‐25), 50 (Ch‐50) or 75 (Ch‐75) mg/day for 12 consecutive weeks. Semen samples were weekly collected from 6th to 9th week of experiment to evaluate some sperm quality parameters including total and progressive motility, plasma membrane integrity and functionality (in fresh and post‐thawed samples) and mitochondrial activity (only in post‐thawed samples). Also, collected semen samples from 10th, 11th and 12th week of experiment were frozen and then artificially inseminated to test fertility rate. According to the results, an improvement in both fresh and post‐thawed sperm quality including total [fresh: 88.00 ± 0.58 and 87.25 ± 0.67 (p < .01); post‐thawed: 51.07 ± 2.05 and 52.72 ± 1.96 (p < .01)] and progressive motility [fresh: 76.00 ± 0.58 and 78.25 ± 0.65 (p < .01); post‐thawed: 40.61 ± 2.01 and 39.88 ± 2.01 (p < .01)], plasma membrane integrity [fresh: 91.60 ± 0.58 and 89.85 ± 0.59 (p < .01); post‐thawed: 56.99 ± 1.86 and 54.39 ± 1.86 (p < .01)] and functionality [fresh: 75.40 ± 0.42 and 77.90 ± 0.96 (p < .01); post‐thawed: 45.69 ± 1.71 and 46.35 ± 1.71 (p < .01)] was noted for both Ch‐50 and Ch‐75, respectively, groups compared to control group. Despite no significant change in mitochondrial activity, fertility rate of post‐thawed spermatozoa was significantly improved in all Chrysin‐fed groups compared to Ch‐0 group. In conclusion, oral Chrysin administration to roosters could ameliorate cryopreservation‐induced impairment of sperm quality and fertility rate.     

Detection of intracellular reactive oxygen species (superoxide anion and hydrogen peroxide) and lipid peroxidation during cryopreservation of alpaca spermatozoa

The objective of this study was to detect changes in intracellular reactive oxygen species (superoxide anion and hydrogen peroxide) production and lipid peroxidation during cryopreservation of alpaca spermatozoa. Twelve alpaca semen samples were conventionally cryopreserved. Intracellular superoxide anion and hydrogen peroxide were evaluated by fluorescence microscopy using dihydroethidium (DHE)/YO‐PRO‐1 and dichlorofluorescein diacetate (H2DCFDA)/propidium iodide (PI), respectively. Evaluations were performed during cooling curve at (1) 25°C, (2) 15°C, (3) 5°C/0 min, (4) 5°C/15 min, (5) 5°C/30 min and (6) after freezing/thawing. Evaluation of lipid peroxidation by measuring malondialdehyde (MDA) was performed at 25°C, 5°C/30 min and after thawing. Maximum percentages of total spermatozoa producing superoxide anion and hydrogen peroxide were found at 5°C/30 min (62.8 ± 6.3% and 30.5 ± 5.6%, respectively), and these results were higher (p < .05) than initial (25°C: 10.8 ± 3.8% and 6.8 ± 0.7%, respectively) and after thawing (29.8 ± 9.5% and 7.5 ± 1.8%, respectively) values. However, considering only viable spermatozoa, production of superoxide anion and hydrogen peroxide during overall stabilization at 5°C (>76% and >91%, respectively) and after thawing (74.9 ± 5.0% and 78.9 ± 2.2%, respectively) was higher (p < .05) than initial values at 25°C (38.7 ± 3.1% and 53.6 ± 2.0%, respectively). Lipid peroxidation at 25°C, 5°C/30 min, and post‐thawing were 346.5 ± 99.8, 401.1 ± 64.8 and 527.7 ± 142.8 ng/ml MDA, respectively. These results showed that high percentage of viable alpaca spermatozoa produces intracellular reactive species oxygen (ROS) during the cryopreservation process of alpaca semen.     

Morphology and quantification of sheep pineal glands at pre‐pubertal,pubertal and post‐pubertal periods

The pineal gland is a neuroendocrine organ associated with photoperiodic regulation in mammals. The aim of this study was to evaluate the pineal gland at the pre‐pubertal, pubertal and post‐pubertal periods by means of morphology and stereology. The study examined at total of 24 ovine pineal glands collected from healthy female Akkaraman breed. Thick sections (40 μm) were cut and treated with synaptophysin. Following each thick section, six consecutive sections at a thickness of 5 μm were cut. Each thin section was stained with one of the following dyes: Crossman’s modified triple dye, glial fibrillary acidic protein (GFAP), melatonin marker, periodic acid–Schiff, Von Kossa and AgNOR. The pineal gland volume was measured using Cavalieri’s method. The optical fractionator was used to estimate the total number of pinealocytes. The percentage of parenchyma and connective tissue and degree of vascularization were estimated by the area fraction fractionator method. The pineal gland volumes in the pre‐pubertal, pubertal and post‐pubertal groups were 7.53 ± 1.715 mm³, 11.20 ± 1.336 mm³ and 17.75 ± 1.188 mm³, respectively (p < .5). The number of pinealocytes in the pre‐pubertal, pubertal and post‐pubertal groups was 3,244,000 ± 228,076, 4,438,000 ± 243,610, 7,381,766 ± 406,223, respectively (p < .05). The glands of the post‐pubertal group contained the highest amount of connective tissue (11.49 ± 2.103%; p < .5) and the largest GFAP staining area (p < .05). The melatonin staining density was the highest in the pubertal group. The density of lipofuscin staining was higher in the pubertal and post‐pubertal groups.     

Alpha‐Linolenic Acid Supplementation in Tris Extender Can Improve Frozen–Thawed Bull Semen Quality

The study was conducted to evaluate the effects of α ‐linolenic acid (ALA) on frozen–thawed quality and fatty acid composition of bull sperm. For that, twenty‐four ejaculates obtained from three bulls were diluted in a Tris extender containing 0 (control), 3, 5, 10 and 15 ng/ml of ALA. Extended semen was incubated at 37°C for 15 min, to allow absorption of ALA by sperm cell membrane. The sample was chilled for 2 h, packed into 0.25‐ml straws and frozen in liquid nitrogen for 24 h. Subsequently, straws were thawed and evaluated for total sperm motility (computer‐assisted semen analysis), membrane functional integrity (hypo‐osmotic swelling test), viability (eosin‐nigrosin), fatty acid composition (gas chromatography) and lipid peroxidation (thiobarbituric acid‐reactive substances (TBARS)). A higher (p < 0.05) percentage of total sperm motility was observed in ALA groups 5 ng/ml (47.74 ± 07) and 10 ng/ml (44.90 ± 0.7) in comparison with control (34.53 ± 3.0), 3 ng/ml (34.40 ± 2.6) and 15 ng/ml (34.60 ± 2.9). Still, the 5 ng/ml ALA group presented a higher (p < 0.05) percentage of viable sperms (74.13 ± 0.8) and sperms with intact membrane (74.46 ± 09) than all other experimental groups. ALA concentration and lipid peroxidation in post‐thawed sperm was higher in all treated groups when compared to the control group. As such, the addition of 5 ng/ml of ALA to Tris extender improved quality of frozen–thawed bull spermatozoa.     

Effect of resveratrol on growth performance,rectal temperature and serum parameters of yellow‐feather broilers under heat stress

This study investigated the effect of dietary resveratrol supplementation on growth performance, rectal temperature, and serum parameters of yellow‐feather broilers under heat stress. A total of 480 yellow‐feather broilers (28‐day‐old) were randomly allotted to five groups with six replicates. A thermoneutral group (TN) (24 ± 2°C) received a basal diet and another four heat‐stressed groups (37 ± 2°C for 8 hr/day and 24 ± 2°C for the remaining time) were fed the basal diet or basal diet with 200, 350, and 500 mg/kg resveratrol for 14 consecutive days. The results revealed that resveratrol supplementation improved average daily gain (p = 0.001), and decreased (p < 0.05) rectal temperature from d 3 when compared with heat‐stressed group without resveratrol. In addition, supplementation with resveratrol at 350 or 500 mg/kg lowered (p < 0.05) the contents of corticosterone, adrenocorticotropic hormone, cholesterol, triglycerides, uric acid, malonaldehyde, and activities of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase, increased (p < 0.05) the levels of triiodothyronine, the ratio of triiodothyronine to thyroxine, total protein, glutathione, and activities of alkaline phosphatase, total superoxide dismutase, catalase, and glutathione peroxidase, though with few fluctuation. In conclusion, supplementation with resveratrol can improve the growth performance by positively regulating serum metabolic parameters and alleviating tissue oxidant damage of broilers under heat stress.     

Evaluation of a point‐of‐care blood glucose monitor in healthy goats

Objective

To assess agreement between a point‐of‐care glucometer (POCG) and a laboratory chemistry analyzer for blood glucose measurements in goats.

Design

Prospective study.

Setting

University teaching hospital.

Animals

Eighteen healthy adult goats.

Investigations

Whole blood samples were obtained via jugular venipuncture prior to premedication with xylazine and butorphanol (T0), following premedication (T20), and after 1 hour of inhalant anesthesia (T60). Each sample was tested with a POCG and a laboratory analyzer (HITA). Agreement was assessed using concordance correlation coefficients and calculation of bias and 95% limits of agreement.

Measurements and Main Results

Mean blood glucose concentration at T0 was 3.9 ± 0.6 mmol/L (70 ± 10 mg/dL; POCG) and 2.9 ± 0.4 mmol/dL (53 ± 8 mg/dL; HITA). Glucose concentrations at T20 were 6.7 ± 2.4 mmol/L (121 ± 43 mg/dL) and 5.4 ± 2.1 mmol/L (97 ± 37 mg/dL) and at T60 were 5.7 ± 1.7 mmol/L (102 ± 31 mg/dL) and 4.7 ± 1.3 mmol/L (85 ± 24 mg/dL) when measured with the POCG and HITA, respectively. The POCG overestimated blood glucose compared to the HITA. The bias ± SD was 1.08 ± 0.53 mmol/L (19.4 ± 9.5 mg/dL) (95% LOA 0.04 to 2.11 mmol/L [0.7 to 38.0 mg/dL]) and the concordance correlation coefficient was 0.82. After correcting the results of the POCG using a mixed‐effects linear model, the bias was 0.0 ± 0.38 mmol/L (0.0 ± 6.8 mg/dL) (95% LOA ± 0.74 mmol/L [± 13.4 mg/dL]) and the concordance correlation coefficient was 0.98.

Conclusions

The POCG overestimated blood glucose concentrations in goats, compared to the HITA, but when the POCG concentrations were corrected, the agreement was excellent.     

Influence of Post‐Mortem Sperm Recovery Method and Extender on Unstored and Refrigerated Rooster Sperm Variables

Many post‐mortem sperm collection techniques have been described for mammalian species, but their use in birds is scarce. This paper compares the efficacy of two post‐mortem sperm retrieval techniques ‐ the flushing and float‐out methods ‐ in the collection of rooster sperm, in conjunction with the use of two extenders, i.e., L&R‐84 medium and Lake 7.1 medium. To determine whether the protective effects of these extenders against refrigeration are different for post‐mortem and ejaculated sperm, pooled ejaculated samples (procured via the massage technique) were also diluted in the above extenders. Post‐mortem and ejaculated sperm variables were assessed immediately at room temperature (0 h), and after refrigeration at 5°C for 24 and 48 h. The flushing method retrieved more sperm than the float‐out method (596.5 ± 75.4 million sperm vs 341.0 ± 87.6 million sperm; p < 0.05); indeed, the number retrieved by the former method was similar to that obtained by massage‐induced ejaculation (630.3 ± 78.2 million sperm). For sperm collected by all methods, the L&R‐84 medium provided an advantage in terms of sperm motility variables at 0 h. In the refrigerated sperm samples, however, the Lake 7.1 medium was associated with higher percentages of viable sperm, and had a greater protective effect (p < 0.05) with respect to most motility variables. In conclusion, the flushing method is recommended for collecting sperm from dead birds. If this sperm needs to be refrigerated at 5°C until analysis, Lake 7.1 medium is recommended as an extender.     

Incorporation of sunflower oil or linseed oil in equine compound feedstuff: 1 Effects on haematology and on fatty acids profiles in the red blood cells membranes

Eight trained horses (6 mares – 2 geldings, 6 Selle Français, 2 Trotteur Français, 12 ± 5.8 years old, 538 ± 72.5 kg) were offered three diets to potentially affect haematology and the fatty acids (FA) profiles in red blood cells (RBC) membranes. The control diet was composed of 50% hay and 50% concentrate containing mainly rolled barley (48%) and whole spelt (48%). In the case of sunflower oil diet, sunflower oil (62.0% of α‐linoleic acid, LA) was incorporated at a rate of 8% and substituted by an equal proportion of barley. In the linseed oil diet, first cold‐pressed linseed oil (56.0% of α‐linolenic acid, ALA) was utilised at a similar incorporation rate of 8%. The experimental design consisted of three 3 × 3 latin squares with one being incomplete. Each period lasted 8 weeks. On average, the total feed intake (straw excluded) was 6.2 kg/day and the oil intake 0.278 kg/day. The oils significantly increased the concentrations of RBC, haemoglobin and haematocrit. The oils had no significant impact on the haematology profiles except that platelets tended to decrease in both oil‐based diets. The most abundant FA in the RBC membranes of the control diet samples were in the decreasing order LA, C18:1n9‐7, C18:0, C16:0 and the arachidonic acid (ARA) respectively. The sunflower oil supplementation slightly increased the amount of LA (36.23 vs. 34.72 mg/dl, p = 0.55) and C22:4n‐6 (0.21 vs. 0.09 mg/dl, p = 0.22), while the decrease was observed in case of other FA (C16:1n‐7, 1.08 vs. 1.42 mg/dl, p = 0.03), C20:3n‐6 (0.22 vs. 0.31 mg/dl, p = 0.02), and ARA (1.17 vs. 1.63 mg/dl, p = 0.08). Linseed oil induced similar effects in the n‐6 series FA profiles. In the context of practical applications, our results show that linseed oil incorporation in the diet could improve the haematology and the n‐3 FA profiles potentially leading to an increased performance.     

Associations between endotoxin‐induced metabolic changes and temperament in Brahman bulls,

The influence of temperament on the alteration of metabolic parameters in response to a lipopolysaccharide (LPS) challenge was investigated. Brahman bulls were selected based on temperament score. Bulls (10 months; 211 ± 5 kg BW; n = 6, 8 and 7 for Calm, Intermediate and Temperamental groups, respectively) were fitted with indwelling jugular catheters to evaluate peripheral blood concentrations of glucose, blood urea nitrogen (BUN), non‐esterified fatty acids (NEFA), insulin, epinephrine and cortisol before and after LPS administration (0.5 μg/kg BW LPS). Feed intake was also recorded. Intermediate bulls consumed more feed than the Temperamental bulls during the challenge (p = 0.046). Pre‐LPS glucose (p = 0.401) and BUN (p = 0.222) did not differ among the temperament groups. However, pre‐LPS insulin (p = 0.023) was lower, whereas pre‐LPS NEFA (p < 0.001), cortisol (p < 0.001) and epinephrine (p < 0.001) were greater in Temperamental than in Calm and Intermediate bulls. Post‐LPS glucose was increased in Calm and Intermediate bulls but not in Temperamental bulls (p < 0.001). Insulin concentrations post‐LPS were greater in Calm than in Intermediate and Temperamental bulls (p < 0.001). Concentrations of NEFA post‐LPS were greater in Temperamental than in Calm and Intermediate bulls (p < 0.001). Serum BUN concentration increased post‐LPS, with values being greater in Calm and Intermediate than in Temperamental bulls (p = 0.012). Collectively, these data demonstrate that animal temperament is related to the metabolic responses of Brahman bulls following a provocative endotoxin challenge. Specifically, Temperamental bulls may preferentially utilize an alternate energy source (i.e. NEFA) to a greater degree than do bulls of Calm and Intermediate temperaments. The use of circulating NEFA from lipolysis may reduce the negative metabolic consequences of an immune response by allowing for a prompt answer to increasing energy demands required during immunological challenge, compared with the time required for glycogenolysis and gluconeogenesis.     

Pharmacokinetic parameters for single‐ and multi‐dose regimens for subcutaneous administration of a high‐dose ceftiofur crystalline‐free acid to neonatal foals

The objective of this study was to determine the pharmacokinetics of single‐ and multi‐dose ceftiofur crystalline‐free acid (CCFA) administered subcutaneously at a dose of 13.2 mg/kg to 12 neonatal foals 1–3 days of age. Six foals received a single subcutaneous dose, while 6 additional foals received 4 doses of CCFA at 48‐h intervals. Blood samples were collected at pre‐determined times following drug administration, and plasma concentrations of ceftiofur free acid equivalents (CFAE) were measured using high‐performance liquid chromatography. Following single‐dose administration of CCFA, the mean ± standard deviation maximum observed plasma concentration was 3.1 ± 0.6 μg/mL and observed time to maximal plasma concentration was 14.0 ± 4.9 h. Following multi‐dose administration of CCFA, the mean ±standard deviation times above CFAE concentrations of ≥0.5 μg/mL and ≥2.0 μg/mL were 192.95 ± 15.86 h and 78.80 ± 15.31 h, respectively. The mean ± standard deviation area under the concentration vs time curve (AUC_0→∝) was 246.2 ± 30.7 h × μg/mL and 172.7 ± 27.14 h × μg/mL following single‐ and multi‐dose CCFA administrations, respectively. Subcutaneous administration of CCFA at 13.2 mg/kg in neonatal foals was clinically well‐ tolerated and resulted in plasma concentrations sufficient for the treatment of most bacterial pathogens associated with neonatal foal septicemia. Multi‐dose administration of four doses at dosing interval of 48 h between treatments maintains appropriate therapeutic concentrations in neonatal foals.     

A retrospective study of female reproductive parameters in the Kunming dog

The Kunming dog is the first and only working dog breed from China to be recognized worldwide. As a domestic working dog, its excellent working performance has been well established; however, its normal reproductive parameters are not well understood. Therefore, this study was conducted to document the main reproductive parameters of this purebred working dog in field breeding conditions. Data on 1004 heats (753 with mating) from 203 bitches between 2008 to 2014, were collected and analyzed. The pregnancy rate and whelping rate was 79.42% and 75.30%, respectively. Finally, for 567 litters (4298 puppies), the mean litter size was 7.19 ± 0.12 puppies (range 1–15). The mean gestation period and birth weight were approximately 61.64 ± 0.10 days and 407.25 ± 1.21 g. The mean sex ratio was 1.03 males to 1.00 female. Estrus occurred throughout the year with no significant differences between seasons and months (P > 0.05), which confirms that Kunming dogs are non‐seasonal breeders; births occurred in every month of the year. Pregnant bitches exhibited significantly longer inter‐estrus intervals than non‐pregnant bitches (220.85 ± 2.05 vs. 180.19 ± 2.94 days, P < 0.05). Bitch parity influenced litter size, and the gestation length and birth weight of the puppies were negatively affected by litter size. This study helps elucidate the reproductive potential of this breed and provides reference values for reproductive performance in the Kunming dog.     

The effect of some cryoprotectants on dromedary camel frozen‐thawed semen

The cryopreserved camel semen is often associated with poor quality and fertility. This study aimed to improve the dromedary frozen semen quality by comparing the efficiency of four cryoprotectant agents (CPAs) on sperm freezability. Semen samples were collected from seven male Maghrabi camels, diluted with Shotor diluent supplemented with glycerol (Sh‐G), dimethyl formamide (DMF, Sh‐DF), dimethyl sulfoxide (DMSO, Sh‐DS) or ethylene glycol (EG, Sh‐EG), all at 6% final concentration, and the samples were subjected to cryopreservation. The results revealed the superiority of Sh‐DF over Sh‐G and Sh‐DS in terms of post‐thaw motility (55.83 ± 2.20 vs. 47.50 ± 4.33 and 45.00 ± 2.89%, respectively), sperm membrane (49.00 ± 0.58, 39.33 ± 3.33 and 42.67 ± 1.45%, respectively) and acrosomal integrities (53.00 ± 0.58, 57.33 ± 0.88 and 52.33 ± 1.45%, respectively). Sh‐EG group showed the lowest post‐thaw motility, plasma membrane and acrosome integrities (12.50 ± 1.44, 22.67 ± 1.45 and 30.67 ± 1.45, respectively). In conclusion, the protocols of dromedary camel semen cryopreservation could be enhanced using 6% DMF as a cryoprotectant agent.