The interleukin 10 response in ovine Johne's disease

Johne's disease is an enteric mycobacterial infection of ruminants that has significant global economic impact. The classic host reaction is one of an early T-cell mediate immune response, with predominant interferon gamma (IFNγ) activity; there is subsequent lowering of this response as animals reach the terminal stages of disease. Interleukin (IL)-10, which can suppress Th1-type and enhance Th2-type cytokine production, is considered to play a role in the later stages of Johne's disease. To determine the role of IL-10 throughout the course of Johne's disease we studied groups of sheep with either no Johne's disease (n = 10), natural infection (n = 30) or experimental infection (n = 58). Disease status of the animals was comprehensively assessed by culture of Mycobacterium avium subsp. paratuberculosis (Mptb), histopathology and serology. Antigen-specific IL-10 secretion in peripheral blood of sheep exposed to Mptb was significantly higher than in control animals (P < 0.001) as early as 4 months post-inoculation, and increased progressively. In ileal and jejunal lymph node cells, IL-10 secretion was also significantly higher in animals that were exposed to Mptb compared to controls (P < 0.05). The early IL-10 response seen in peripheral blood cells may be a reflection of early responses at sites of Mptb infection. IL-10 secretion from ileal and jejunal lymph node cells was significantly higher in exposed animals with no lesions or with paucibacillary lesions when compared to animals with multibacillary lesions. These novel findings demonstrate that increased IL-10 activity commences soon after exposure to the causative mycobacterium and may play a role in determining disease outcome.     

Measurement of vitamin A,vitamin E,selenium, and L-lactate in dogs with and without osteoarthritis secondary to ruptured cranial cruciate ligament

This study compared vitamin A, vitamin E, selenium (Se), and L-lactate in blood and synovial fluid in 2 groups of 6 dogs; a control group (without OA) and an osteoarthritic group with spontaneous cranial cruciate ligament rupture and OA. Concentrations of vitamin E were significantly higher in serum than in synovial fluid in both OA (P = 0.006) and control (P = 0.0008) groups. Vitamin E concentration in synovial fluid was significantly higher in the OA group than in the control group (P = 0.009). Concentrations of Se were significantly higher in serum than in synovial fluid in both OA (P = 0.003) and control (P = 0.0006) groups. There were no significant differences in levels of Se, vitamin A, and L-lactate between the 2 groups. This is the first study to show an increased concentration of vitamin E in the synovial fluid of dogs with OA compared with dogs that did not have OA.     

The Neospora caninum-Spain 7 isolate induces placental damage, fetal death and abortion in cattle when inoculated in early gestation

The Nc-Spain 7 isolate of Neospora caninum, which was newly obtained from an asymptomatic congenitally infected calf, demonstrated a similar virulence as Nc-1 strain in mouse models. The aim of this study was to characterize the pathogenesis of Nc-Spain 7 isolate in cattle after experimental infection at 65 days of gestation. For this purpose, thirteen pregnant heifers were divided into three groups as follows: group A: 7 heifers inoculated with 1×10(8) tachyzoites of Nc-Spain 7 isolate; group B: 4 heifers inoculated with 1×10(8) tachyzoites of Nc 1 strain; and group C: 2 heifers received PBS. Serum samples were collected weekly and heparinized blood samples were collected three times (0, 28 and 42 days after inoculation) by jugular venipuncture. Placenta and fetal tissue samples were collected at time of necropsy. Specific antibody response in the dams was tested by IFAT, indirect ELISA, and rNcGRA7 and rNcSAG4 based-ELISA. Specific antibody response in fetal fluids was tested by IFAT. IFN-γ production was measured after in vitro culture of PBMC and the supernatant was assessed using a commercial kit (BOVIGAM). A significant increase in N. caninum antibody responses was detected in groups A and B by IFAT and by i-ELISA from day 14 after inoculation onwards. Besides, antibody response against rNCGra7 protein was also detected in all inoculated heifers by rNcGra7-based ELISA. Four fetuses from group A and one from group B were aborted between 3 and 5 weeks after infection. In the recovered fetuses, only 3 out of 4 fetal fluids from fetuses of group A and 1 out of 3 of group B were seropositive by IFAT, but all of them were positive by PCR. Transplacental transmission could be determined in all fetuses from groups A and B by PCR and/or IHC. Heifers of group C and their fetuses remained negative by all techniques. The results of this study demonstrate that the NC-Spain 7 isolate could be transmitted transplacentally, and produced fetal death and abortion in cattle.     

Effect of Progesterone and Ionomycin on Domestic Cat Sperm Motility Patterns and Acrosome Reaction

This study was aimed at assessing the changes in sperm motion patterns and the percentage of acrosome reaction (AR) in domestic cat semen after treatment with either ionomycin or progesterone (P₄). Ten ejaculates were collected from five tomcats using an artificial vagina, and were diluted, centrifuged and resuspended in a capacitation medium. Samples were evaluated and divided into seven equal aliquots and, after 2 h at 25°C, were incubated for 30 min at 38°C in 5% CO₂ and then analyzed. Computer-assisted sperm analysis and a combination of three fluorescent probes were used to assess sperm plasma, acrosomal membrane integrity and mitochondrial transmembrane potential. Thirty minutes after the start of incubation, P₄ was added (10 μg/ml) to the P1 group. Groups P2 and P3 were supplemented with P₄ (10 and 20 μg/ml, respectively) only after 2 h of incubation, and groups I1 and I2 were supplemented with ionomycin (4 and 8 μ m , respectively) 2 h after incubation. Group E was supplemented with ethanol (0.6%) at 2 h after incubation and group C received no supplementation. Ionomycin and P₄ treatments led to a hyperactivation-like sperm motion and an increase (p < 0.05) in the percentage of AR. Although a higher (p < 0.05) percentage of AR was obtained in group I2 when compared with all P₄ groups, a decrease (p < 0.05) in total and progressive motility was observed in I2 group. As I1 group was similar to I2 to induce AR without diminishing sperm motility, we can conclude that ionomycin at 4 μ m seems to be more suitable to trigger AR in domestic cat sperm.     

The pharmacokinetics and metabolism of ivermectin in domestic animal species

The pharmacokinetic properties of drugs are closely related to their pharmacological efficacy. The kinetics of ivermectin are characterised, in general terms, by a slow absorption process, a broad distribution in the organism, low metabolism, and slow excretion. The kinetics vary according to the route of administration, formulation, animal species, body condition, age, and physiological status, all of which contribute to differences in drug efficacy. Characterisation of ivermectin kinetics can be used to predict and optimise the value of the parasiticide effects and to design programmes for parasite control. This article reviews the pharmacokinetics of ivermectin in several domestic animal species.     

Technical note: recombinant LHRH fusion protein suppresses estrus in heifers

A recombinant luteinizing hormone-releasing hormone (LHRH) fusion protein was evaluated for its effectiveness in suppression of estrus in heifers. Eight heifers were randomly assigned to two equal treatment groups. Treatments consisted of recombinant ovalbumin-LHRH-7 or recombinant ovalbumin (control). This recombinant chimeric fusion protein consisted of ovalbumin with seven LHRH peptides (ovalbumin-LHRH-7). The plasmid for this protein was expressed in E. coli and was collected and purified as an insoluble protein. One milligram of the respective proteins was suspended in 2 mL of Z-Max adjuvant and administered by intramammary injection three times at 7-wk intervals. Luteinizing hormone-releasing hormone antibody binding was elevated in heifers treated with ovalbumin-LHRH-7 compared to ovalbumin-treated heifers (P < .05). Serum progesterone concentrations (< 1 ng/mL) indicate that the estrous cycle of the four heifers treated with ovalbumin-LHRH-7 was suppressed for a time period ranging from 60 to 238 d, which was different from control heifers (P < .01). Serum progesterone for the control heifers continued to exhibit cyclic profiles over the experimental period. This preliminary study in heifers demonstrated that a chimeric LHRH fusion protein induced elevated concentrations of circulating LHRH antibodies that suppressed estrus for an average of 122 +/- 41 d.     

Use of whole blood for spectrophotometric determination of cholinesterase activity in dogs

Whole blood has been compared with erythrocytes and plasma for spectrophotometric cholinesterase determination in the dog. Cholinesterase activity was characterized using two substrates: acetylthiocholine and butyrylthiocholine. Acetylcholinesterase was the only form of cholinesterase present on erythrocytes and hydrolysed only acetylthiocholine. Butyrylcholinesterase (pseudocholinesterase) was predominant in plasma, hydrolysing mainly butyrylthiocholine. Based on these results, a method based on the use of two substrates (acetylthiocholine for monitoring acetylcholinesterase and butyrylthiocholine for determining butyrylcholinesterase) in the same whole blood sample is recommended for canine cholinesterase analysis. This way of monitoring both enzymes can be easily automated, yielding good within (CVs < 5%) and between-run (CVs < 7%) precision.     

Influence of storage time and processing on chemical composition and in vitro ruminal fermentation of olive cake

Olive oil extraction generates olive cake (OC) that could be used in ruminant feeding. However, the chemical composition of OC is affected by multiple factors, being therefore highly variable. The objective of this study was to analyse the influence of storage time and further processing: crude, exhausted (subjected to a second oil extraction) and cyclone (obtained from a cyclone separator) on nutritive value of OC samples. Twelve samples (six crude and six exhausted) were obtained monthly from the same pond from 1 to 6 storage months, and nine samples (three crude, three exhausted and three cyclone) were obtained monthly from a different pond from 6 to 9 months storage. Chemical composition was analysed, and OC samples were fermented in vitro with sheep rumen fluid. Increasing storage time up to 6 months decreased sugars and total soluble polyphenols content but increased fibre content in OC. Dry matter effective degradability (DMED) decreased linearly (p < 0.001) by 35.9 and 45.5% as storage time augmented from 1 to 6 months for crude and exhausted OC, respectively. Crude OC had lower DMED values than exhausted OC (averaged values 0.255 and 0.294 g/g, respectively). Both potential production and rate of gas production were lower (p ≤ 0.018) in crude compared with exhausted OC, which was attributed to the high fat content of crude OC (≥86 g/kg dry matter). For samples stored longer than 6 months, cyclone had greater (p < 0.05) DMED than crude and exhausted OC (averaged values 0.207, 0.164 and 0.164 g/g, respectively). The results indicate that ruminal degradability of OC is reduced with advancing storage time, but only subtle changes were observed during the first two months. Cyclone showed greater degradability than crude and exhausted OC, but differences between crude and exhausted OC became negligible after five storage months.     

Dexamethasone and prednisolone in the horse: pharmacokinetics and action on the adrenal gland

Pharmacokinetics of dexamethasone and prednisolone were studied in 6 horses given dexamethasone alcohol (IV or IM) or dexamethasone 21-isonicotinate as a solution IV or IM (50 micrograms/kg of body weight), prednisolone 21-sodium succinate IV or IM (0.6 mg/kg of body weight), or prednisolone acetate IM (0.6 mg/kg of body weight). Plasma concentrations were determined using a high-performance liquid chromatographic method. After dexamethasone alcohol (IV) or dexamethasone 21-isonicotinate (IV), the half-life of elimination was similar (53 minutes) for both formulations. After dexamethasone (alcohol and isonicotinate, IM), concentrations were low or nondetected. After prednisolone 21-sodium succinate (IV), the half-life of elimination (99.5 minutes) was significantly (P less than 0.01) longer than that for dexamethasone. After prednisolone 21-sodium succinate (IM), absorption was rapid and bioavailability was high. After prednisolone acetate (IM), absorption was slow and prednisolone was present in plasma for about 7 days. Due to the nonlinearity of prednisolone kinetics, a bioavailability higher than 100% was obtained. The basal plasma hydrocortisone concentration was approximately 70 ng/ml. After dexamethasone (IV or IM), plasma hydrocortisone values decreased after a 2-hour delay and returned to base line after a 3 to 4 day delay. After prednisolone 21-sodium succinate (IV or IM), plasma hydrocortisone decreased immediately (IV) or rapidly (IM) and returned to base line after a 24-hour delay. After prednisolone acetate (IM), plasma hydrocortisone decreased for up to 21 days.