Jugular Vein Blood Samples Research Articles

Assessment of oxidative stress parameters in summer anestrous buffaloes exhibiting differential fertility following melatonin implants treatment

For evaluating the impact of melatonin implants treatment during non-breeding season to ameliorate oxidative stress, 132 anestrous buffaloes were subcutaneously inserted with 2x4 mm absorbable slow-release melatonin implants (18 mg/50 kg b wt) at the base of left ear and 60 buffaloes were used as control. Ovarian ultrasonography and jugular vein blood sampling were carried out at 7-day interval till day 35 post-treatment or till ovulation, whichever was earlier. Control and implanted buffaloes were subjected to artificial insemination (AI) at overt or induced estrus followed by pregnancy diagnosis at day 90 post-AI. Erythrocytic lipid peroxidation (LPO) values were reduced (P<0.05) in implanted buffaloes from day 21 post-treatment onwards when compared to their pre-treatment and Control group values. However, the concentrations of erythrocytic glutathione peroxidase (GPx), glutathione reductase and superoxide dismutase (SOD) were invariably higher (P<0.05) following treatment as compared to their pre-treatment and Control group values. The buffaloes ovulating in Control or Treatment group revealed higher (P<0.05) erythrocytic GPx in the Latter group. Also, between pregnant counterparts of Control and Treatment group, the Latter group buffaloes exhibited low (P<0.05) erythrocytic LPO, and high (P<0.05) erythrocytic GPx, SOD and catalase. It can be concluded that melatonin implants treatment was successful for mitigating the oxidative stress in summer anestrous buffaloes, and the status of oxidative stress parameters following treatment was better in buffaloes that ovulated or conceived subsequently.

29 The Role of Leptin and Its Regulation in the Hypothalamus of Neonatal Calves

Abstract The constant occurrence of increased feed intake in offspring exposed to intrauterine growth restriction has been noted across multiple species including ruminants. In ruminants, reduced circulating leptin concentrations from birth through the first week or two of life may influence the development of the appetite center of the hypothalamus. It should able be noted that increased circulating cortisol at birth has been associated with this decreased circulating leptin in the perinatal ruminants, yet information is limited as to any cause and effect relationship between the two hormones. The objectives of these studies were to test directly the effects of exogenous cortisol on leptin concentrations relative to development of the hypothalamus in perinatal calves and voluntary feed intake as yearling. In two experiments, Holstein bulls (n = 27) and calves (n = 32 males; n = 30 females) from commercial Angus cows were assigned to treatments at parturition (day 0). Calves were intravenously infused with either a low cortisol [LC; n = 9, 3.5 ug/kg of body weight (BW)], high cortisol (HC; n = 9, 7.0 ug/kg BW), or a control (CON; n = 9, saline). Calves were administered a half dose of their respective treatment 24h postpartum. Jugular vein blood samples were collected before infusion and daily until d 17 or necropsy. At 5 days of age Holstein bull calves were euthanized where cerebral-spinal fluid (CSF) and hypothalamic tissue were collected. Serum and CSF leptin concentrations were analyzed via RIA. Neuronal growth factors were analyzed in hypothalamus samples via qRT-PCR. In experiment 2, yearling calves entered a GROWSAFE System and daily feed intake (FI) measured. Animals were allowed a two-week adjustment period to a growing ration for heifers and a finishing ration for steers. Heifer body condition scores (BCS) were collected at the beginning and end of the trial. Heifers underwent the feeding trial for 70 days and steers until they obtained a back fat (BF) thickness of 1.15 cm. Blood samples were analyzed similar to experiment 1. All data were analyzed via repeated measures using appropriate models of SAS (SAS Institute Inc., Cary, NC). Perinatal dairy bull calves had decreased (P < 0.013) serum and CSF leptin concentrations of HC and LC calves compared with CON. Hypothalamic expression of BDNF, FGF1 and FGF2 were decreased (P < 0.006) in HC and LC compared with CON. During the feeding trial: BW gain, BCS change, and number of feed events were increased (P = 0.001) in LC compared with HC and CON heifers. However, LC observed greater daily FI (P = 0.047) and tended to have greater final BW (P = 0.080) compared with HC and CON steers. In summary, these data support that exogenous cortisol administered to perinatal calves affects leptin production and the appetite center of the hypothalamus when administered during the neonatal period.

Assessment of plasma melatonin profile in summer anestrous buffaloes exhibiting differential fertility following melatonin implants treatment

Plasma melatonin vis-a-vis anestrus or estrus status of buffaloes during summer or winter season (n=50 in each group) suggested its impact on reproductive axis as plasma melatonin activity tended to be higher (P>0.05) in buffaloes exhibiting estrus compared to their anestrous counterparts irrespective of season. The differential reproductive status of these buffaloes was confirmed by history as well as differences (P<0.05) in diameter of largest follicle and luteal profile. Further, for evaluating the impact of melatonin treatment during summer season on reproductive axis, 132 anestrous buffaloes were subcutaneously inserted 2x4 mm absorbable slow-release melatonin implants (18 mg/50 kg b wt) at the base of left ear and 60 buffaloes were used as control. In these buffaloes, ovarian ultrasonography and jugular vein blood sampling was carried out at 7-day interval till day 35 post-treatment or till ovulation, whichever was earlier. Control and implanted buffaloes were subjected to artificial insemination (AI) at overt or induced estrus followed by pregnancy diagnosis at day 90 post-AI. In treatment group, an increase (P<0.05) in plasma melatonin was recorded in all the buffaloes, exhibiting differential fertility status, during post-treatment study period compared to their pre-treatment and control group values. However, within treatment group, there was no difference (P>0.05) in plasma melatonin between ovulatory or non-ovulatory as well as between pregnant or non-pregnant counterparts. Moreover, plasma melatonin within control buffaloes remained similar (P>0.05) throughout the study period irrespective of differential exhibition of fertility. It can be concluded that factors other than circulating melatonin are also involved in the display of differential fertility in terms of initiation of ovarian cyclicity/ovulation or ability to conceive in summer anestrous buffaloes.

Serological Detection, Isolation and Molecular Confirmation of Parainfluenza Virus-3 in Camels, Iraq

The objectives of this study were to detect and isolate the Parainfluenza-3 virus (PIV-3) in camels with naturally developed respiratory illness and to determine the titer of the isolates using the virus titration. Therefore, an overall 100 nasal swabs and jugular vein blood samples were collected from diseased camels in four districts in Wasit province (Iraq) from December (2019) to March (2020). The swabs were subjected to six subsequent passages on bovine kidney cell culture (BKCC) to isolate the virus and to confirm infection by molecular PCR assay. Fever (40°C), abundant runny nasal discharge, ocular discharge, coughing, depression, increased respiratory rate, abnormal breath sounds, and mainly wheezing are the most observed clinical signs. Positive findings were involved 24% by ELISA and 37% by RT-PCR. The age group from 1-2 years old showed a high infection rate, while the lower level was in the 4-6 years old group. Regarding season, the infection rate was high in winter compared to spring. Sheik Saad city appeared to have a higher infection rate than other districts. The positive samples inoculated into the Bovine kidney cell culture (BKCC) revealed the cytopathic effects (CPE) after three successive passages, which appeared as clumping and rounding with the progression of infection time at the 4th passage. Elongation and giant cell formation were shown in some isolates after the 5th and 6th passages until they reached complete detachments of the cells from the cell sheet. The titer of viral tissue culture infective dose (TCID50) of the 3rd passage was determined in BKCC cells at 10–3/0.05 ml, and the high titer was shown at the 5th and the 6th passages equal to 10-5/ 0.05 ml. In conclusion, PIV-3 is widespread among camels infected with respiratory illness; therefore, studies are necessary to detect the prevalence rate among camels in other Iraqi regions. Keywords: PIV-3, Fusion protein gene, Hemagglutination protein gene, ELISA, PCR

Increasing the prepartum dose of rumen-protected choline: Effects of maternal choline supplementation on growth, feed efficiency, and metabolism in Holstein and Holstein × Angus calves

Feeding pregnant cows rumen-protected choline (RPC) may have the potential to affect the growth and health of offspring, but little is known about the optimal dose, or the potential mechanisms of action. The objectives of this experiment were to 1) determine if increasing RPC supplementation during late gestation in multiparous Holstein cows would improve calf growth and 2) determine if maternal choline supplementation alters global DNA methylation patterns. Pregnant multiparous Holstein cows (n = 116) were randomly assigned to diets targeting 0g choline ion (0.0 ± 0.000 choline ion, %DM, control; CTL), 15g of choline ion (recommended dose; RD) from an established RPC product (0.10 ± 0.004 choline ion, %DM, RPC1RD; ReaShure, Balchem Corp.; positive control), or 15g (0.09 ± 0.004 choline ion, %DM, RPC2RD) or 22g (0.13 ± 0.005 choline ion, %DM, high dose; RPC2HD) of choline ion from a concentrated RPC prototype (RPC2; Balchem Corp.). Treatments were mixed into a total mixed ration and cows had ad libitum access via a roughage intake control system (Hokofarm Group, Marknesse, Netherlands). All female Holstein (n = 49) and Holstein × Angus calves (male, n = 18; female, n = 30) were enrolled and fed colostrum from a cow within the same treatment. Holstein calves and Holstein × Angus calves were fed an accelerated and traditional milk replacer program, respectively, and offered ad libitum access to calf starter. Jugular vein blood samples were collected, and body weight was measured at 7, 14, 28, 42, and 56 d of age. Categorical treatment and continuous effects of actual prepartum maternal choline ion intake were analyzed using mixed effect models. An interaction of treatment with sex, nested within breed, resulted in any choline treatment increasing the proportion of methylated whole blood DNA in male, but not female calves. Although 37% of Holstein calves across all treatments experienced abomasal bloat, no evidence for differences in health measurements (signs of respiratory disease and fecal consistency) were observed across treatments. During the first 2 wk of life in Holstein calves, RPC2HD tended to increase average daily gain (ADG) and feed efficiency (FE) compared with CTL and increasing maternal choline ion intake linearly increased ADG and FE. Maternal choline supplementation increased plasma glucose compared with CTL, while increasing serum insulin-like growth factor-1 and decreasing serum lipopolysaccharide binding protein at 7 d of age in Holstein calves. In Holstein × Angus calves, the effect of treatment on ADG tended to interact with sex: in males, RPC2HD increased ADG after 2 wk of life compared with CTL, without evidence of a treatment effect in female calves. Increasing maternal choline ion intake linearly increased ADG after 2 wk of age in male Holstein × Angus calves, while quadratically increasing FE in both sexes. Altered global DNA methylation patterns in male Holstein × Angus calves, and changes in blood metabolites in Holstein calves, provide 2 potential mechanisms for observed improvements in calf growth. Continuous treatment models demonstrated that the effects of maternal choline supplementation are sensitive to the amount of maternal choline ion intake, with greater benefit to calves observed at higher maternal intakes.

Effect of melatonin implantation prior to ovsynch plus cidr protocol on subsequent ovarian activity and pregnancy rate in summer anestrous Murrah buffaloes

For assessing the impact of melatonin pre-treatment in summer anestrous Murrah buffaloes on subsequent reproductive activity of a fixed time artificial insemination (FTAI) protocol, 83 anestrous Murrah buffaloes were subcutaneously inserted 2x4 mm size absorbable melatonin (MT) implants (18 mg/50 kg b wt) at the base of left ear and 38 Murrah buffaloes were used as non-implanted controls. On day 35 post-MT, the treatment and control group Murrah buffaloes were subjected to FTAI protocol (Ovsynch plus CIDR). Ovarian ultrasonography and jugular vein blood sampling was carried out on day 7 and 10 of protocol and on day 7-post AI. Overall pregnancy rate on day 90 post-AI in MT group was higher compared to control group (55.4 vs 31.6%, P<0.05), along with the higher (P>0.05) plasma progesterone on day 7 of FTAI protocol in Murrah buffaloes of the former group which conceived subsequently compared to their non-pregnant counterparts. Moreover, MT group had higher (P>0.05) increase in dominant follicle diameter between day 7 and 10 of protocol compared to control group. On day 7 post-AI, the luteal profile in pregnant murrah buffaloes of MT group was better (P<0.05) compared to their counterparts in control group (CL: 14.80±0.70 vs 10.82±0.46 mm, P4: 1.54±0.07 vs 1.04±0.08 ng/ml). In conclusion, melatonin pre-treatment in summer anestrous Murrah buffaloes could be associated with better luteal activity and higher pregnancy rate in the Ovsynch+CIDR protocol.

Essential amino acid profile of supplemental metabolizable protein affects mammary gland metabolism and whole-body glucose kinetics in dairy cattle.

This study investigated mammary gland metabolism and whole-body (WB) rate of appearance (Ra) of glucose in dairy cattle in response to a constant supplemental level of metabolizable protein (MP) composed of different essential AA (EAA) profiles. Five multiparous rumen-fistulated Holstein-Friesian dairy cows (2.8 ± 0.4 lactations; 81 ± 11 d in milk; mean ± standard deviation) were abomasally infused according to a 5 × 5 Latin square design with saline (SAL) or 562 g/d of EAA delivered in different profiles where individual AA content corresponded to their relative content in casein. The profiles consisted of (1) a complete EAA mixture (EAAC), (2) Ile, Leu, and Val (ILV), (3) His, Ile, Leu, Met, Phe, Trp, Val (GR1+ILV), and (4) Arg, His, Lys, Met, Phe, Thr, Trp (GR1+ALT). A total mixed ration (58% corn silage, 16% alfalfa hay, and 26% concentrate on a dry matter basis) was formulated to meet 100 and 83% of net energy and MP requirements, respectively, and was fed at 90% of ad libitum intake on an individual cow basis. Each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of no infusion. Arterial and venous blood samples were collected on d 4 of each period for determination of mammary gland AA and glucose metabolism. On d 5 of each period, D-[U-13C]glucose (13 mmol priming dose; continuous 3.5 mmol/h for 520 min) was infused into a jugular vein and arterial blood samples were collected before and during infusion to determine WB Ra of glucose. Milk protein yield did not differ between EAAC, GR1+ILV, and GR1+ALT, or between SAL and ILV, and increased over SAL and ILV with EAAC and GR1+ILV. Mammary plasma flow increased with ILV infusion compared with EAAC and GR1+ILV. Infusion of EAAC tended to increase mammary gland net uptake of total EAA and decreased the mammary uptake to milk protein output ratio (U:O) of non-EAA compared with SAL. Infusion of ILV increased mammary net uptake and U:O of Ile, Leu, and Val markedly over all treatments. The U:O of total Ile, Leu, and Val increased numerically (25%) with GR1+ILV infusion compared with EAAC, and the U:O of total Arg, Lys, and Thr tended to decrease, primarily from decreased U:O of Lys. During GR1+ALT infusion, U:O of total Arg, Lys, and Thr was greater than that during EAAC infusion, whereas U:O of Ile, Leu, and Val did not differ from EAAC. Glucose WB Ra increased 16% with GR1+ALT over SAL, and increased numerically 8 and 12% over SAL with EAAC and GR1+ILV, respectively. The average proportion of lactose yield relative to glucose WB Ra did not differ across treatments and averaged 0.53. On average, 28% of milk galactose arose from nonglucose precursors, regardless of treatment. In conclusion, intramammary catabolism of group 2 AA increased to support milk component synthesis when the EAA profile of MP was incomplete with respect to casein. Further, WB and mammary gland glucose metabolism was flexible in support of milk component synthesis, regardless of absorptive EAA profile.

Physicochemical Analysis of Mixed Venous and Arterial Blood Acid-Base State in Horses at Core Temperature during and after Moderate-Intensity Exercise.

Simple SummaryThe purposes of the present study were to determine the effect of core body temperature on acid-base variables and determine the origins of acid-base changes in the arterial and mixed venous blood of horses during exercise and recovery. Moderate intensity exercise resulted in an increase in body temperature that, in addition to exercise, affected acid-base status and gas partial pressures. Moderate intensity exercise resulted in a mild alkalosis that had markedly different origins in arterial blood than in mixed venous blood, and this was affected by the increase in core temperature during exercise and its resolution during recovery. In order to fully understand how acid-base status changes during exercise and recovery, it is importance to quantify the changes in both arterial and mixed venous blood, with adjustment to core temperature. Acid-base assessments using jugular vein blood samples are limited in comparison.The present study determined the independent contributions of temperature, strong ion difference ([SID]), total weak acid concentration ([Atot]) and PCO2 to changes in arterial and mixed venous [H+] and total carbon dioxide concentration ([TCO2]) during 37 min of moderate intensity exercise (~50% of heart rate max) and the first 60 min of recovery. Six horses were fitted with indwelling carotid and pulmonary artery (PA) catheters, had PA temperature measured, and had blood samples withdrawn for immediate analysis of plasma ion and gas concentrations. The increase in core temperature during exercise (+4.5 °C; p < 0.001) significantly (p < 0.05) increased PO2, PCO2, and [H+], but without a significant effect on [TCO2] (p > 0.01). The physicochemical acid-base approach was used to determine contributions of independent variables (except temperature) to the changes in [H+] and [TCO2]. In both arterial and venous blood, there was no acidosis during exercise and recovery despite significant (p < 0.05) increases in [lactate] and in venous PCO2. In arterial blood plasma, a mild alkalosis with exercise was due to primarily to a decrease in PCO2 (p < 0.05) and an increase in [SID] (p < 0.1). In venous blood plasma, a near absence of change in [H+] was due to the acidifying effects of increased PCO2 (p < 0.01) being offset by the alkalizing effects of increased [SID] (p < 0.05). The effect of temperature on PO2 (p < 0.001) resulted in an increased arterio-venous PO2 difference (p < 0.001) that would facilitate O2 transfer to contracting muscle. The simultaneous changes in the PCO2 and the concentrations of the other independent acid-base variables (contributions from individual strong and weak ions as manifest in [SID] and [Atot]) show complex, multilevel control of acid-base states in horses performing even moderate intensity exercise. Correction of acid-base variables to core body temperature presents a markedly different physiological response to exercise than that provided by variables measured and presented at an instrument temperature of 37 °C.

Differences in hormone levels around parturition in Hanwoo cattle (Bos taurus coreanae) following artificial insemination and embryo transfer.

BackgroundWith unique genetic traits, Hanwoo cattle (Bos taurus coreanae) are well‐adapted to the Korean environment. However, their perinatal mortality rate is 2%–3%, which imposes an economic burden.ObjectiveDue to insufficient data on hormonal changes around parturition, the timing of parturition is often predicted subjectively; few studies have examined hormones in Hanwoo cattle. We measured the changes in various hormones around parturition, to seek an objective predictor of parturition time.MethodsIn 14 female Hanwoo cattle, we measured progesterone, prolactin and cortisol concentrations daily in jugular vein blood samples, beginning 6 days before parturition until 7 days after parturition. Conception was induced in five animals using artificial insemination. Nine animals received embryo transfer.ResultsDuring parturition, the concentration of progesterone decreased significantly in the embryo transfer group (n = 9) and in the total population (n = 14); it did not change significantly in the artificial insemination group (n = 5). Prolactin concentration increased on the day of parturition but did not differ significantly among the groups. Cortisol remained constant throughout the study course.ConclusionWe concluded that parturition time can be predicted in Hanwoo cattle using progesterone concentration. This knowledge can reduce perinatal mortality, which would help to improve farm income and animal welfare.

Agreement between arterial and central venous blood pH and its contributing variables in anaesthetized dogs with respiratory acidosis

ObjectiveTo determine the accuracy of variables that influence blood pH, obtained from central venous (jugular vein) blood samples compared with arterial (dorsal pedal artery) samples in anaesthetized dogs with respiratory acidosis. Study designProspective, comparative, observational study. AnimalsA group of 15 adult male dogs of various breeds weighing 17 (11-42) kg [median (range)]. MethodsDogs were premedicated with buprenorphine (0.03 mg kg–1) and medetomidine (0.01 mg kg–1) administered intramuscularly by separate injections, anaesthetized with propofol intravenously to effect and maintained with isoflurane in 50% air-oxygen. Arterial and central venous catheters were placed. After 15 minutes of spontaneous breathing, arterial and central venous blood samples were obtained and analysed within 5 minutes, using a bench-top gas analyser. Differences between arterial and central venous pH and measured variables were assessed using Wilcoxon rank sum test and effect size (r: matched-pairs rank-biserial correlation) was calculated for each comparison. The agreement (bias and limits of agreement: LoAs) between arterial and central venous pH and measured variables were assessed using Bland-Altman; p < 0.05. Data are reported as median and 95% confidence interval. ResultsArterial blood pH was 7.23 (7.19-7.25), and it was significantly greater than central venous samples 7.21 (7.18-7.22; r = 0.41). Agreement between arterial and venous pH was acceptable with a bias of 0.01 (0.002-0.02) and narrow LoAs. PCO2 [arterial 54 (53-58) mmHg, 7.2 (7.1-7.7) kPa; venous 57 (54-62) mmHg, 7.6 (7.2-8.3) kPa], bicarbonate ion concentration and base excess did not differ between samples; however, agreement between arterial and venous PCO2 was not acceptable with a bias of –2 (–5 to 0) mmHg and wide LoAs. Conclusions and clinical relevanceBlood pH measured from central venous (jugular vein) blood is an acceptable clinical alternative to arterial blood (dorsal pedal artery) in normovolaemic anaesthetized dogs with respiratory acidosis.

Factors associated with Anaplasma phagocytophilum infection in sheep in Iran

Anaplasma phagocytophilum is one of the most widespread tick-borne pathogens in sheep across the world. The aim of this cross-sectional study was the determination of the molecular prevalence and associated risk factors of A. phagocytophilum infection in sheep populations of Iran. For this aim, between March 2018 and February 2019 jugular vein blood samples were collected from 1782 apparently healthy sheep from nine provinces from four ecological zones of Iran. Specific nested-PCR targeting the 16S rRNA gene of A. phagocytophilum was employed. Twelve variables were subjected to logistic regression analyses (univariate and multivariate) to specify the potential risk factors for infection. Statistically significant variables in univariate analyses (P ≤ 0.20) were assessed by multivariable logistic regression to control the confounding factors. DNA of Anaplasma phagocytophilum was detected in 8.1 % of animals (144/1782). Among geographical zones, the prevalence was highest in the Persian Gulf zone (P<0.001); and among provinces Mazandaran (in the north) had the highest prevalence (P < 0.001). The multivariate analysis revealed that no/irregular application of ectoparasiticides (OR=2.09, P < 0.001), short-distance between farms (less than 1km) (OR=1.96, P = 0.011), and high-density barns (<1.5 m2 /head) (OR = 1.45, P = 0.050) are potential risk factors of A. phagocytophilum infection in sheep from Iran. The spatial scan statistic in SaTScan recognized three high risks clusters for A. phagocytophilum infection in Caspian (Mazandaran Province) and the Persian Gulf (Khuzestan Province) zones amongst the study areas (P < 0.001). Phylogenetic analysis based on A. phagocytophilum 16S rRNA genes revealed significant nucleotide sequence identities between the sequences obtained in this study with those previously collected from other countries. This research is the largest study focusing on ovine anaplasmosis in Iran and shows that infected sheep are present in all zones, and geographic areas. The connection between A. phagocytophilum and its transmission routes in the country e.g. ticks, iatrogenesis, and economic impact of the infection remains largely unknown and warrants further investigation.

Anaplasma ovis infection in sheep from Iran: molecular prevalence, associated risk factors, and spatial clustering.

The aim of this cross-sectional study was to determine the molecular prevalence and associated risk factors in sheep populations of Iran. To this end, between March 2017 and February 2018 jugular vein blood samples were collected from 1842 apparently healthy sheep from 327 herds in nine provinces in four ecological zones of Iran. A specific nested-PCR targeting the msp4 gene of A. ovis was employed. Fourteen variables were subjected to logistic regression analyses (univariate and multivariate) to specify the potential risk factors for infection. Statistically significant variables in univariate analyses (P ≤ 0.20) were assessed by multivariable logistic regression to control the confounding factors. Anaplasma ovis DNA was detected in 51.1% of herds (167/327) and 28.3% of animals (521/1842). Among geographical zones, herd and animal prevalence was highest in the Persian-Gulf zone (P < 0.001), and among provinces, Lorestan (in west) and Khuzestan (in south-west) had the highest prevalence (P < 0.001). Analysis of factors associated with A. ovis infection revealed that distance from other farms (OR = 2.52, P < 0.001), presence of other animal species in the farm (OR = 2.03, P = 0.046), season (OR = 1.40, P = 0.005), breed (OR = 3.762, P < 0.001), and age of sheep (OR = 1.20, P = 0.049) are potential risks in Iran. The spatial scan statistic inSaTScan recognized two high risks clusters for A. ovis infection in central (Semnan province) and the Persian-Gulf (Khuzestan province) zones amongst the study areas (P < 0.001). Sequence and phylogenetic analysis of the msp4 gene confirmed the detection of A. ovis. This research is the largest study focusing on ovine anaplasmosis in Iran and shows that infected sheep are present in all geographic zones, bioclimatic areas, and provinces.

Lung-protective ventilation increases cerebral metabolism and non-inflammatory brain injury in porcine experimental sepsis

BackgroundProtective ventilation with lower tidal volumes reduces systemic and organ-specific inflammation. In sepsis-induced encephalopathy or acute brain injury the use of protective ventilation has not been widely investigated (experimentally or clinically). We hypothesized that protective ventilation would attenuate cerebral inflammation in a porcine endotoxemic sepsis model. The aim of the study was to study the effect of tidal volume on cerebral inflammatory response, cerebral metabolism and brain injury. Nine animals received protective mechanical ventilation with a tidal volume of 6 mL × kg−1 and nine animals were ventilated with a tidal volume of 10 mL × kg−1. During a 6-h experiment, the pigs received an endotoxin intravenous infusion of 0.25 µg × kg−1 × h−1. Systemic, superior sagittal sinus and jugular vein blood samples were analysed for inflammatory cytokines and S100B. Intracranial pressure, brain tissue oxygenation and brain microdialysis were sampled every hour.ResultsNo differences in systemic or sagittal sinus levels of TNF-α or IL-6 were seen between the groups. The low tidal volume group had increased cerebral blood flow (p < 0.001) and cerebral oxygen delivery (p < 0.001), lower cerebral vascular resistance (p < 0.05), higher cerebral metabolic rate (p < 0.05) along with higher cerebral glucose consumption (p < 0.05) and lactate production (p < 0.05). Moreover, low tidal volume ventilation increased the levels of glutamate (p < 0.01), glycerol (p < 0.05) and showed a trend towards higher lactate to pyruvate ratio (p = 0.08) in cerebral microdialysate as well as higher levels of S-100B (p < 0.05) in jugular venous plasma compared with medium–high tidal volume ventilation.ConclusionsContrary to the hypothesis, protective ventilation did not affect inflammatory cytokines. The low tidal volume group had increased cerebral blood flow, cerebral oxygen delivery and cerebral metabolism together with increased levels of markers of brain injury compared with medium–high tidal volume ventilation.

Puerarin enhances intestinal function in piglets infected with porcine epidemic diarrhea virus

Puerarin has been reported to be an excellent antioxidant, anti-inflammatory and antimicrobial agent, but the potential effect of puerarin on porcine epidemic diarrhea virus (PEDV) is unclear. This study aimed to determine whether puerarin could alleviate intestinal injury in piglets infected with PEDV. A PEDV (Yunnan province strain) infection model was applied to 7-day-old piglets at 104.5 TCID50 (50% tissue culture infectious dose). Piglets were orally administered with puerarin at the dosage of 0.5 mg/kg body weight from day 5 to day 9. On day 9 of the trial, piglets were inoculated orally with PEDV. Three days later, jugular vein blood and intestinal samples were collected. Results showed puerarin reduced morbidity of piglets infected with PEDV. In addition, puerarin reduced the activities of aspartate aminotransferase and alkaline phosphatase, the ratio of serum aspartate aminotransferase to serum alanine aminotransferase, the number of white blood cells and neutrophils, and the plasma concentrations of interleukin-6, interleukin-8 and tumor necrosis factor-α, as well as protein abundances of heat shock protein-70 in PEDV-infected piglets. Moreover, puerarin increased D-xylose concentration but decreased intestinal fatty acid-binding protein concentration and diamine oxidase activity in the plasma of piglets infected with PEDV. Puerarin increased the activities of total superoxide dismutase, glutathione peroxidase and catalase, while decreasing the activities of myeloperoxidase and concentration of hydrogen peroxide in both the intestine and plasma of PEDV-infected piglets. Puerarin decreased mRNA levels of glutathione S-transferase omega 2 but increased the levels of nuclear factor erythroid 2-related factor 2. Furthermore, puerarin increased the abundance of total eubacteria (16S rRNA), Enterococcus genus, Lactobacillus genus and Enterobacteriaceae family in the intestine, but reduced the abundance of Clostridium coccoides in the caecum. These data indicate puerarin improved intestinal function in piglets infected by PEDV and may be a promising supplement for the prevention of PEDV infection.

Mechanism of CuSO4 cytotoxicity in goat erythrocytes after high-level in vitro exposure to isotonic media

Copper (Cu) is a common environmental pollutant in nature. Cu-poisoning can cause liver damage and erythrocytes hemolysis. To evaluate the effect of CuSO4 poisoning on the morphological and functional characteristics of goat red blood cells. Five 10–14-month-old goats were selected for jugular vein blood sampling to obtain erythrocytes, and then the erythrocytes were processed with different concentrations (0, 10, 20, 30, 40 and 50 μmol/L) of CuSO4 for 48 h, and 40 μmol/L doses CuSO4 incubated for different time (12, 24, 36, 48 and 60 h) to process erythrocytes. We observed the changes in erythrocyte morphology through scanning electron microscopy, and detected the antioxidant function and activities of three ATPases. Additionally, biological properties were examined from the perspectives of phospholipids and membrane protein components, permeability fragility, and fluidity in erythrocytes. We found that after CuSO4 treatment, the antioxidant capacity of erythrocytes decreased, which was manifested as increased MDA content and decreased CuZn-SOD and GSH-Px activities (p < 0.05). In addition, we also found that erythrocyte fluidity decreased, osmotic fragility increased, membrane phospholipid percentage and protein composition changes abnormally, and Na+/K+-ATPase, Mg2+-ATPase and Ca2+-ATPase activities decreased (p < 0.05). From the results, it can be concluded that CuSO4 exposure causes hemolysis of goat erythrocytes through oxidative stress to the structure and function of erythrocytes, showing a dose-time effect.

PSIX-25 Porcine epidemic diarrhea virus infection impairs intestinal nutrient transport in piglets

Abstract Porcine epidemic diarrhea virus (PEDV) infection can occur in all ages of pigs, but neonatal piglets are the most susceptible and sensitive to the virus. PEDV infection can cause intestinal dysfunction, severe diarrhea and even death in piglets. This study determined the effects of PEDV infection on the absorptive function and gene expression of nutrient transporters in the small intestine of piglets by microarray assay. Sixteen 7-day-old healthy piglets fed with milk replacer and were randomly allocated to one of two treatments (the Control and PEDV groups). After a 5-day adaptation period, piglets (n = 8) were orally administrated with either sterile saline or PEDV (Yunnan province strain) solution at 104.5TCID50 (50% tissue culture infectious dose) per piglet. On day 5 of the trial, D-xylose (0.1 g/kg BW) was orally administrated to all piglets. One hour later, jugular vein blood samples were collected, and then all piglets were killed to obtain the intestinal samples. Compared with the control, PEDV infection increased (P &amp;lt; 0.05) incidence of diarrhea, plasma DAO activity and iFABP level, while decreased (P &amp;lt; 0.05) plasma D-xylose concentration of piglets. Moreover, PEDV infection altered the amino acids profiles (P &amp;lt; 0.05), and decreased (P &amp;lt; 0.05) the gene expression of AQPs (AQP4, AQP8 and AQP10), amino acids transporters (y+LAT1, b0’+AT, and PepT1), molecules associated with lipid transport and metabolism (LPL, SLC27A2, and ACSL3), and glucose transport and metabolism (GLUT2 and INSR). However, PEDV infection enhanced (P &amp;lt; 0.05) the gene epression of PCK1, ASS1, SGLT1, and CFTR in the jejunum of piglets. Collectively, these comprehensive results indicate that PEDV infection impairs intestinal absorptive function and inhibits the expression of genes associated with nutrient transport and metabolism in piglets.

Microarray analysis reveals the inhibition of intestinal expression of nutrient transporters in piglets infected with porcine epidemic diarrhea virus

Porcine epidemic diarrhea virus (PEDV) infection can induce intestinal dysfunction, resulting in severe diarrhea and even death, but the mode of action underlying these viral effects remains unclear. This study determined the effects of PEDV infection on intestinal absorption and the expression of genes for nutrient transporters via biochemical tests and microarray analysis. Sixteen 7-day-old healthy piglets fed a milk replacer were randomly allocated to one of two groups. After 5-day adaption, piglets (n = 8/group) were orally administrated with either sterile saline or PEDV (the strain from Yunnan province) at 104.5 TCID50 (50% tissue culture infectious dose) per pig. All pigs were orally infused D-xylose (0.1 g/kg BW) on day 5 post PEDV or saline administration. One hour later, jugular vein blood samples as well as intestinal samples were collected for further analysis. In comparison with the control group, PEDV infection increased diarrhea incidence, blood diamine oxidase activity, and iFABP level, while reducing growth and plasma D-xylose concentration in piglets. Moreover, PEDV infection altered plasma and jejunal amino acid profiles, and decreased the expression of aquaporins and amino acid transporters (L-type amino acid transporter 1, sodium-independent amino acid transporter, B(°,+)-type amino acid transport protein, sodium-dependent neutral amino acid transporter 1, sodium-dependent glutamate/aspartate transporter 3, and peptide transporter (1), lipid transport and metabolism-related genes (lipoprotein lipase, apolipoprotein A1, apolipoprotein A4, apolipoprotein C2, solute carrier family 27 member 2, solute carrier family 27 member 4, fatty acid synthase, and long-chain acyl-CoA synthetase (3), and glucose transport genes (glucose transporter-2 and insulin receptor) in the jejunum. However, PEDV administration increased mRNA levels for phosphoenolpyruvate carboxykinase 1, argininosuccinate synthase 1, sodium/glucose co-transporter-1, and cystic fibrosis transmembrane conductance regulator in the jejunum. Collectively, these comprehensive results indicate that PEDV infection induces intestinal injury and inhibits the expression of genes encoding for nutrient transporters.

Cyclical cervical function in the mare involves remodelling of collagen content, which is correlated with modification of oestrogen receptor 1 abundance

This study was conducted to elucidate mare cervical dilation mechanisms by testing two hypotheses: (i) the proportion of collagen staining in histological samples of mare cervices and (ii) the abundance of hormone receptors in the equine cervix differ with stage of the oestrous cycle and site within the cervix. Tissues and jugular vein blood samples were collected from 15 mares. Collagen content was assessed using Masson’s Trichome staining. Receptor abundance was assessed using RT-PCR, qRT-PCR and immunohistochemistry. In sub-epithelial stroma, there was less collagen during the follicular than luteal phase, in the caudal- (P = 0.029), mid- (P = 0.0000) and cranial (P = 0.001) cervical tissue. In the deep stroma, there was less collagen staining during the follicular stage in the mid- (P = 0.004) and cranial- (P = 0.041) cervical regions. There were PTGER2, PTGER3, PGR and ESR1 mRNA transcripts in the cervix. A greater proportion of cells were positive for ESR1 protein during the follicular phase in sub-epithelial (P = 0.019) and deep (P = 0.013) stroma. The abundance of ESR1 in the epithelium was negatively correlated with collagen staining in sub-epithelial (P = 0.007) and deep (P = 0.005) stroma. The results of the study provide new information about the cervical biology of mares by increasing the knowledge about collagen content and the relationship between collagen content and ESR1 protein abundance during the oestrous cycle which indicates the ESR1 receptor is a candidate for involvement in control of cervical dilation.

Effects of prostaglandin F2α on angiogenic and steroidogenic pathways in the bovine corpus luteum may depend on its route of administration

Prostaglandin (PG) F2α and its analogs (aPGF2α) are used to induce regression of the corpus luteum (CL); their administration during the middle stage of the estrous cycle causes luteolysis in cattle. However, the bovine CL is resistant to the luteolytic actions of aPGF2α in the early stage of the estrous cycle. The mechanisms underlying this differential luteal sensitivity, as well as acquisition of luteolytic sensitivity by the CL, are still not fully understood. Therefore, to characterize possible differences in response to aPGF2α administration, we aimed to determine changes in expression of genes related to (1) angiogenesis-fibroblast growth factor 2 (FGF2), fibroblast growth factor receptor 1 (FGFR1), fibroblast growth factor receptor 2 (FGFR2), vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2); and (2) steroidogenesis-steroidogenic acute regulatory protein (STAR), cytochrome P450 family 11 subfamily A member 1 (P450scc), and hydroxy-delta-5-steroid dehydrogenase, 3 β- and steroid delta-isomerase 1 (HSD3B) in early- and middle-stage CL that accompany local (intra-CL) versus systemic (i.m.) aPGF2α injection. Cows at d 4 (early stage) or d 10 (middle stage) of the estrous cycle were treated as follows: (1) systemic saline injection, (2) systemic aPGF2α injection (25 mg), (3) local saline injection, and (4) local aPGF2α injection (2.5 mg). Progesterone (P4) concentration was measured in jugular vein blood samples during the entire set of experiments. After 4 h of treatment, CL were collected by ovariectomy, and mRNA and protein expression levels were determined by reverse transcription quantitative-PCR and Western blotting, respectively. Local and systemic aPGF2α injections upregulated FGF2 expression but decreased expression of VEGFA in both CL stages. Both aPGF2α injections increased the expression of STAR in early-stage CL, but downregulated it in middle-stage CL. In the early-stage CL, local administration of aPGF2α upregulated HSD3B, whereas systemic injection decreased its mRNA expression in early- and middle-stage CL. Moreover, we observed a decrease in the P4 level earlier after local aPGF2α injection than after systemic administration. These results indicate that aPGF2α acting locally may play a luteotrophic role in early-stage CL. The systemic effect of aPGF2α on the mRNA expression of genes participating in steroidogenesis seems to be more substantial than its local effect in middle-stage CL.

Influence of unilateral ovariectomy performed before ovulation on ovarian function, steroid hormone levels and development of porcine fetuses

Corpus luteum (CL) activity is closely linked with initiation and maintenance of pregnancy and with fetal development. The present study was aimed to analyze the impact of unilateral ovarian removal on ovarian function, steroid hormone level and fetal distribution and development. Unilateral ovariectomy (uni-OVX) was performed in gilts one day before ovulation (group SHORT, n = 24), 20 days before subsequent ovulation (group LONG, n = 23) or ovaries remained intact (group INTACT, n = 22). Gilts were inseminated by single fixed-time laparoscopic intrauterine insemination (LIUI) after hormonal estrus synchronization. Two days before the end of a 15 day long altrenogest feeding, a part of gilts (n = 23) were surgically fitted with a jugular vein catheter and blood samples were collected to determine of estradiol (E2) and progesterone (P4) concentrations. All animals were slaughtered on day 30 of gestation and ovarian features as well as the number, weight and distribution of fetuses recorded. Altogether, 48 gilts (70%) were pregnant and pregnancy rates did not differ between groups. Short term uni-OVX affected CL number compared to intact and long term OVX gilts (9.9 ± 0.8 vs. 20.6 ± 1.9 and 17.5 ± 0.8; P < 0.05) and the number of fetuses (8.7 ± 0.5 vs. 15.0 ± 1.0 and 14.4 ± 1.1; P<0.05), respectively. Weights of individual CL were not influenced by treatment. Fetuses were differently distributed in the uterine horns after uni-OVX. A higher (P<0.05) proportion of fetuses was present always in the horn which bore the ovary. In all groups, weights of fetuses from uterine horns with an active ovary was similar; however, fetuses of the OVX horn of the SHORT group were lighter (P < 0.05). Steroid hormone profile was typical for pregnant gilts, but differences were observed between groups. Both, the preovulatory E2 concentrations and the early luteal phase P4 levels were higher (P < 0.05) in gilts of the INTACT and LONG groups. In summary, compared to intact gilts, only long term uni-OVX could compensate ovarian development. Short term uni-OVX affects (1) the total number of CL and fetuses, (2) the distribution and weight of fetuses in the uterine horns and (3) steroid hormone levels. Therefore, the time window, but not the uni-OXV per se, which alters the local supply of progesterone, has an impact on fetal development and survival.