The use of intrarenal injection of sodium pentobarbital for euthanasia in cats

OBJECTIVE To determine whether intrarenal injection of sodium pentobarbital is a viable method for euthanasia in anesthetized client-owned cats and assess potential factors associated with time to cardiopulmonary arrest (TCPA) for such treated cats. ANIMALS 131 client-owned cats. PROCEDURES In this retrospective study, client-owned cats presented for euthanasia between March 1, 2009, and January 15, 2010, were evaluated by veterinarians to determine suitability of intrarenal injection versus other methods of euthanasia. Cats included were anesthetized and then received 6 mL of sodium pentobarbital (390 mg/mL) by intrarenal injection. Results for TCPA were compared for cats grouped on the basis of variables of interest. RESULTS 131 cats were included, of which 74 (79%) had a TCPA < 1 minute and 28 (21%) had a TCPA between 1.5 and 8 minutes after intrarenal injection. Most (124/131 [95%]) cats had no observable reaction to the intrarenal injection other than cardiopulmonary arrest. Median TCPA was longer for cats without (1 min; 25/131 [19%]) versus with (0 min; 106/131 [81%]) palpable kidney swelling upon injection. CLINICAL RELEVANCE The effects of intrarenal injection of sodium pentobarbital in cats of the present study were similar to those typically observed with IV administration of euthanasia solution. When this intrarenal injection method is used, cardiopulmonary arrest with few agonal reactions can be expected to occur quickly in most patients. The intrarenal injection method is suited for euthanasia of anesthetized cats with easily located kidneys when IV access may be difficult.

Wound healing by topical application of Momordica charantia L. formulations on mice

Background and Aim: Momordica charantia is mainly characterized by its antimicrobial and antioxidant properties. The current study aimed to evaluate the healing activity of gel and cream formulations based on M. charantia on induced wounds in mice. Materials and Methods: Acetonic extract of M. charantia was prepared and incorporated into gel and cream formulations. Mus musculus Balb/c (n=30) with induced injury were distributed into five groups: Group I (control – day 7), Group II (control – day 14), Group III (1% gel – day 7), and Group IV (1% gel – day 14) to which 1% M. charantia gel was dermally applied daily for 7 and 14 days, respectively, Group V (1% cream – day 7) and Group VI (1% cream – day 14) to which of M. charantia 1% cream were dermally applied daily for 7 and 14 days, respectively. Time of wound closure was determined during the experimentation; rats were euthanized with sodium pentobarbital 60 mg/kg/pc v.ip. for obtaining skin samples for histopathological analysis. Results: Groups IV and VI showed a higher percentage of wound closure on day 14, and in histopathological analysis, effect was greater in Group VI with the presence of fibroblasts and abundant collagen and elastic fibers. Conclusion: M. charantia gel and cream showed wound healing activity on induced wounded mice; the most effective treatment was M. charantia 1% cream formulation.

Effects of Exogenous Glucocorticoid Administration on Appetitic Center Brain Development in Dairy Bull Calves

The objective of study was to determine the effects of exogenous cortisol on leptin concentrations and appetitic center development of perinatal dairy bull calves. Holstein bull calves (n = 27) were weighed and randomly assigned to treatments within 4 h of parturition (d0). Each calf was intravenously infused with either a low cortisol (LC; n = 9, 3.5 ug/kg of BW), high cortisol (HC; n = 9, 7.0 ug/kg of BW), or a sham infusion control (CON; n = 9, similar volume of saline). Each calf was administered a second infusion (half dose) of its respective treatment 24 h postpartum. All calves were housed similarly and fed milk replacer (28% CP, 20% fat) three times daily. Blood was collected via jugular venipuncture before infusion and from d 0–5 of age. At 5 d of age, calves were euthanized via sodium pentobarbital overdose. Cerebral-spinal fluid (CSF) from the third ventricle of the brain, hypothalamus, and adipose tissue (AT; omental, perirenal, and mesenteric) were collected. Blood and CSF samples were analyzed for leptin concentrations via a validated RIA. Adipose tissue samples were analyzed via western blotting for leptin and glucocorticoid receptor (GR) expression and normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression. Hypothalamus samples were analyzed via qRT-PCR for genes associated with neuronal growth factors and normalized to the average of two housekeeping genes. All data were analyzed via ANOVA or repeated measures analysis using appropriate models (SAS Institute Inc., Cary, NC). Serum and CSF leptin concentrations were decreased (P &lt; 0.013) in HC and LC compared to CON. Leptin protein expression was decreased (P &lt; 0.044) in perirenal and omental AT of LC calves compared to CON. Hypothalamic expression of BDNF, FGF1, and FGF2 were decreased (P &lt; 0.006) in HC and LC compared to CON. In summary, exogenous cortisol administered to calves at birth reduced leptin concentrations and altered appetitic control center development of the brain.

Rapid activation of esophageal mechanoreceptors alters the pharyngeal phase of swallow: Evidence for inspiratory activity during swallow

Swallow is a complex behavior that consists of three coordinated phases: oral, pharyngeal, and esophageal. Esophageal distension (EDist) has been shown to elicit pharyngeal swallow, but the physiologic characteristics of EDist-induced pharyngeal swallow have not been specifically described. We examined the effect of rapid EDist on oropharyngeal swallow, with and without an oral water stimulus, in spontaneously breathing, sodium pentobarbital anesthetized cats (n = 5). Electromyograms (EMGs) of activity of 8 muscles were used to evaluate swallow: mylohyoid (MyHy), geniohyoid (GeHy), thyrohyoid (ThHy), thyropharyngeus (ThPh), thyroarytenoid (ThAr), cricopharyngeus (upper esophageal sphincter: UES), parasternal (PS), and costal diaphragm (Dia). Swallow was defined as quiescence of the UES with overlapping upper airway activity, and it was analyzed across three stimulus conditions: 1) oropharyngeal water infusion only, 2) rapid esophageal distension (EDist) only, and 3) combined stimuli. Results show a significant effect of stimulus condition on swallow EMG amplitude of the mylohyoid, geniohyoid, thyroarytenoid, diaphragm, and UES muscles. Collectively, we found that, compared to rapid cervical esophageal distension alone, the stimulus condition of rapid distension combined with water infusion is correlated with increased laryngeal adductor and diaphragm swallow-related EMG activity (schluckatmung), and post-swallow UES recruitment. We hypothesize that these effects of upper esophageal distension activate the brainstem swallow network, and function to protect the airway through initiation and/or modulation of a pharyngeal swallow response.

Two Cases of Natural Infection of Dengue-2 Virus in Bats in the Colombian Caribbean

Dengue, a mosquito-borne zoonotic disease, is the most common vector-borne disease in tropical and subtropical areas. In this study, we aim to demonstrate biological evidence of dengue virus infection in bats. A cross-sectional study was carried out in the departments of Cordoba and Sucre, Colombia. A total of 286 bats were captured following the ethical protocols of animal experimentation. The specimens were identified and euthanized using a pharmacological treatment with atropine, acepromazine and sodium pentobarbital. Duplicate samples of brain, heart, lung, spleen, liver, and kidney were collected with one set stored in Trizol and the other stored in 10% buffered formalin for histopathological and immunohistochemical analysis using polyclonal antibodies. Brain samples from lactating mice with an intracranial inoculation of DENV-2 were used as a positive control. As a negative control, lactating mouse brains without inoculation and bats brains negative for RT-PCR were included. Tissue sections from each specimen of bat without conjugate were used as staining control. In a specimen of Carollia perspicillata captured in Ayapel (Cordoba) and Phylostomus discolor captured in San Carlos (Cordoba), dengue virus was detected, and sequences were matched to DENV serotype 2. In bats RT-PCR positive for dengue, lesions compatible with viral infections, and the presence of antigens in tissues were observed. Molecular findings, pathological lesions, and detection of antigens in tissues could demonstrate viral DENV-2 replication and may correspond to natural infection in bats. Additional studies are needed to elucidate the exact role of these species in dengue epidemics.

Effects of dietary fennel (Foeniculum vulgare Mill.) seed powder supplementation on growth performance, nutrient digestibility, small intestinal morphology, and carcass traits of broilers

Background With the increasing demands in livestock and poultry breeding and the growing number of food-borne diseases, it is necessary to practice food safety and develop strategies to produce healthy livestock. Fennel (Foeniculum vulgare Mill.) has been used as an additive in poultry production by some researchers, but there are few studies on the systemic beneficial effects of dietary fennel seed powder supplementation on broilers. Therefore, this study aimed to investigate the effect of dietary fennel seed powder supplementation on feed intake, the apparent metabolic rate of nutrients, intestinal morphology, and carcass traits in Cobb broilers. Methods A single-factor experimental design was used. In total, 160 1-day-old Cobb broiler chicks were randomly assigned to four treatments, with four replicates each (n = 10/replicate). Broilers in the control (CN) group were fed a basal diet without fennel seed powder, and broilers in the treatment groups were fed a basal diet supplemented with 0.15% (LF), 0.30% (MF), or 0.45% (HF) fennel seed powder, respectively. Feeding trials lasted for 42 days under the conditions of ad libitum access to feed and water, and 24-h illumination. During the third and sixth weeks, digestive and metabolic assays were carried out. When the broilers were 42 days old, one chicken with a weight close to the average was selected from each repetition, euthanized by an intravenous injection of 5% sodium pentobarbital, and carcass traits were measured and intestinal samples were collected for morphological assessment. Results There was no significant difference in growth performance of broilers (P > 0.05). The breast muscle percentage, fat width and fat width index, breast muscle area, and breast muscle area index of broilers in the LF group were higher than those in other groups (P < 0.05). Jejunum weight and length were higher in MF than in CN and LF broilers (P < 0.05). Additionally, duodenal villi height, ileal villi height, and ileal wall thickness were higher in MF than in CN broilers (P < 0.05). There were no significant differences in nutrient utilization among all groups (P > 0.05), except that the ash apparent metabolic rate in MF broilers at 21 days of age was higher than that in LF broilers (P < 0.05). In conclusion, dietary supplementation with a moderate concentration of fennel affects carcass performance, and intestinal morphology, and promotes the growth and development of broilers.

Establishment and effect evaluation of an aortic dissection model induced by different doses of β-aminopropionitrile in rats

Introduction Given the controversy regarding the appropriate dose of β-aminopropionitrile for induction of aortic dissection models in rats, the purpose of this study was to explore the most suitable concentration of β-aminopropionitrile to establish a high-incidence and low-mortality aortic dissection model. Methods Eighty three-week-old male Sprague-Dawley rats were equally divided into four groups: a control group, a 0.06% β-aminopropionitrile group, a 0.08% β-aminopropionitrile group and a 0.1% β-aminopropionitrile group. Initial experiments were performed on the control group, which was not treated with β-aminopropionitrile (and drank water freely), and the other three groups, which were given different concentrations of β-aminopropionitrile solution daily (0.06%, 0.08% and 0.1%). Subsequently, on the 40th day, osmotic minipumps administering 1 μg/kg per min angiotensin II (Ang II) were implanted subcutaneously into the β-aminopropionitrile groups, while the control group was continuously pumped with normal saline. The rats were euthanized 48 h after implantation. All rats that died before the expected end time of the experiment were autopsied immediately, and the aortas were dissected. The rats surviving at the end of the experiment were sacrificed by an overdose of sodium pentobarbital, and tissue samples were harvested for further analyses. Results The mean survival days were significantly different among the groups, with 39.1 ± 6.04 days in the 0.08% β-aminopropionitrile group and 32.7 ± 9.85 days in the 0.1% β-aminopropionitrile group ( P = 0.0178) at the end of the experiment. Compared with those in the 0.06% β-aminopropionitrile group, the rates of aortic dissection were significantly higher in the 0.08% β-aminopropionitrile group and the 0.1% β-aminopropionitrile group ( P = 0.0015 and P = 0.0005, respectively), while there was no significant difference between the 0.08% β-aminopropionitrile group and the 0.1% β-aminopropionitrile group ( P = 0.723) at 70% and 75%, respectively. However, the rupture rates were significantly different between the 0.08% β-aminopropionitrile group and the 0.1% β-aminopropionitrile group (55% versus 20%, P = 0.022). Hematoxylin-eosin staining of the aortic tissue sections of the β-aminopropionitrile group showed that red blood cells entered the pseudocavity in the vascular wall, while the vascular wall structure of the control group was intact. Compared with control rats, which were intact and free from fracture, β-aminopropionitrile-treated rats had fewer collagen fibers and exhibited fracture. Magnetic resonance imaging showed that the aortic intimae of the aortic dissection rats showed double lumens and intimal tears. Conclusions An aortic dissection model with a high incidence and low mortality was successfully and stably developed with 0.08% β-aminopropionitrile. This model will enable further studies investigating aortic dissection pathogenesis and drug therapy. Magnetic resonance imaging may be a reliable technique for imaging the aorta in rats.

Lactobacillus salivarius, a Potential Probiotic to Improve the Health of LPS-Challenged Piglet Intestine by Alleviating Inflammation as Well as Oxidative Stress in a Dose-Dependent Manner During Weaning Transition

Intestinal health is a critical issue for piglets during their weaning transition period. Previous reports have emphasized the promise of distinct probiotics in improving the enteric health. Here in this research, a newly isolated Lactobacillus salivarius strain was pretreated to Lipopolysaccharide (LPS)-challenged piglets and its association with integrity of the intestinal barrier coupled with effective dosage were expected to be signified. In the present study, 72 piglets (Landrace × Yorkshiere × Duroc) were randomly allotted to four groups, each group with six replicates. The subjects in the control group were provided with basal diet while those in other tested groups with extra 0.05, 0.1, and 0.2% L. salivarius, respectively. Fourteen days later, LPS was intraperitoneally injected and sodium pentobarbital was then delivered to euthanize those LPS-challenged piglets. An increase of average daily gain and body weight along with an apparent decline of diarrhea rate were observed in L. salivarius-treated groups. Both 0.1 and 0.2% L. salivarius supplement in total diet had the capability to markedly elevate levels of CAT, GSH-Px, SOD, anti-inflammatory cytokine from the serum as well as tight junction proteins (Claudin-1, Occludin, and ZO-1) extracted from intestine in LPS-challenged piglets. These changes were accompanied by the obvious downregulation of D-lactic acid, DAO, MDA and pro-inflammatory mediators in the serum, including IL-1β, IL-6, IFN-γ, and TNF-α. Meanwhile, the expression levels of TLR2 and TLR4 in spleen and mesenteric lymph nodes were significantly lower whereas the oxidation-related gene, ho-1 was up-regulated with L. salivarius administration. Our findings suggested that relatively high dose L. salivarius (0.1–0.2%) could regulate the progression of inflammatory response and oxidative stress when individuals were exposed to LPS, thus probably offering valuable assistance in restoring barrier function and improving overall performance.

60 Effects of exogenous glucocorticoid administration on leptin concentrations in perinatal dairy bull calves

Previous studies suggest a relationship between leptin and cortisol immediately postpartum is linked to improved feedlot efficiency on cattle exposed to perinatal stressors. The objective of the current study was to determine the effects of exogenous cortisol administration on circulating leptin concentrations in perinatal dairy bull calves. Within four hours of parturition, Holstein bull calves were randomly assigned to treatments and intravenously infused with low cortisol (LC; n =9, 3.5 ug/kg of cortisol), high cortisol (HC; n = 9, 7.0 ug/kg of cortisol), or a sham infusion control (CON; n = 9, similar volume of saline). Calves were housed similarly and fed milk replacer (28% CP, 20% fat) three times daily. Blood collections were performed via jugular venipuncture using Z/9mL serum collection tubes (Sarstedt, Newton, NC) from d 0 to 5 of age. Serum samples were incubated for 1 h at room temperature then overnight at 4°C, centrifuged at 1800 x g at 4°C for 20 minutes, serum decanted, and stored long term at -20°C until analysis. At 5 d of age, calves were euthanized utilizing an overdose of sodium pentobarbital (Schering-Plough Animal Health, Union, NJ) and cerebral-spinal fluid (CSF) from the third ventricle of the brain was collected and stored at -80°C until analysis. All samples were analyzed for leptin concentrations via a previously-validated radioimmunoassay. Serum leptin concentrations were analyzed via repeated measures of ANOVA and CSF leptin concentrations via the GLM procedure of SAS (SAS Institute Inc., Cary, NC, USA). Serum leptin concentrations of HC and LC were decreased (P = 0.013) compared to CON, and d 0 was decreased (P &lt; 0.001) compared to all other days of age. Cerebral-spinal fluid leptin concentrations of HC and LC were decreased (P = 0.005) compared to CON. In summary, exogenous cortisol administered to perinatal dairy bull calves reduced leptin concentrations in serum and CSF.

59 Effects of exogenous glucocorticoid administration on leptin concentrations in perinatal dairy bull calves

Previous studies suggest a relationship between leptin and cortisol immediately postpartum is linked to improved feedlot efficiency on cattle exposed to perinatal stressors. The objective of the current study was to determine the effects of exogenous cortisol administration on circulating leptin concentrations in perinatal dairy bull calves. Within four hours of parturition, Holstein bull calves were randomly assigned to treatments and intravenously infused with low cortisol (LC; n = 9, 3.5 ug/kg of cortisol), high cortisol (HC; n = 9, 7.0 ug/kg of cortisol), or a sham infusion control (CON; n = 9, similar volume of saline). Calves were housed similarly and fed milk replacer (28% CP, 20% fat) three times daily. Blood collections were performed via jugular venipuncture using Z/9mL serum collection tubes (Sarstedt, Newton, NC) from d 0 to 5 of age. Serum samples were incubated for 1 h at room temperature then overnight at 4°C, centrifuged at 1800 x g at 4°C for 20 minutes, serum decanted, and stored long term at -20°C until analysis. At 5 d of age, calves were euthanized utilizing an overdose of sodium pentobarbital (Schering-Plough Animal Health, Union, NJ) and cerebral-spinal fluid (CSF) from the third ventricle of the brain was collected and stored at -80°C until analysis. All samples were analyzed for leptin concentrations via a previously-validated radioimmunoassay. Serum leptin concentrations were analyzed via repeated measures of ANOVA and CSF leptin concentrations via the GLM procedure of SAS (SAS Institute Inc., Cary, NC, USA). Serum leptin concentrations of HC and LC were decreased (P = 0.013) compared to CON, and d 0 was decreased (P &lt; 0.001) compared to all other days of age. Cerebral-spinal fluid leptin concentrations of HC and LC were decreased (P = 0.005) compared to CON. In summary, exogenous cortisol administered to perinatal dairy bull calves reduced leptin concentrations in serum and CSF.