Serum Levels of Cardiac Markers NT‐proANP and NT‐proBNP in Brachycephalic bitches at Different Gestational Stages

The aim of this study was to determine serum levels of natriuretic peptide precursors (NT‐proANP and NT‐proBNP) during pregnancy in brachycephalic bitches. Fifteen healthy multiparous bitches were selected for this prospective study. Serum levels of NT‐proANP and NT‐proBNP were measured during anoestrous and at 14, 35, 42, 49 and 56 days (2nd, 5th, 6th, 7th and 8th weeks) of pregnancy. Fourteen animals had normal gestations, and one bitch developed single foetus syndrome. The natriuretic peptide levels of this animal were not included in this study; however, it is important to report that its NT‐proANP levels were four times greater than those of normal patients. There was no significant difference (p = 0.072) in NT‐proBNP levels between anoestrous (0.20 ± 0.10 ng/ml) and the different pregnancy weeks (0.27 ± 0.12 ng/ml). There was a positive correlation (p < 0.0001) between NT‐proANP and gestational age, and the levels of this marker increased significantly (p < 0.0001) during the 6th (0.26 ± 0.06 ng/ml), 7th (0.28 ± 0.04 ng/ml) and 8th weeks (0.29 ± 0.05 ng/ml) when compared to anoestrous (0.18 ± 0.02 ng/ml). NT‐proANP serum levels are correlated with gestational development and may be indicative of cardiovascular adaptation in canine brachycephalic pregnancy.     

Leukocyte emigration in the early stages of laminitis

The mechanisms that initiate the pathophysiologic changes in the digital laminae in equine laminitis are poorly understood. Due to the fact that (1) the horse at risk of laminitis has many similarities clinically to the human sepsis patient and (2) our recent finding of marked laminar proinflammatory cytokine expression at the developmental time point of the black walnut extract (BWE) model of laminitis, we tested the possibility that, similar to organ damage in human sepsis, leukocyte emigration is an early event in laminitis. Using immunoperoxidase methods with an anti-equine CD13 monoclonal antibody that recognizes neutrophils and monocytes, we discovered that, whereas the dermal microvasculature of the skin commonly has a marginal pool of leukocytes, the normal laminar dermal microvasculature has minimal to no perivascular leukocytes. However, increases in leukocyte numbers occurred around the dermal vasculature of both the laminae and the skin in the majority of BWE-treated horses in the developmental stage and at the onset of clinical signs of lameness in the BWE model. These findings indicate that, similar to organ failure in human sepsis, leukocyte emigration is likely to play a significant role in initiating numerous pathophysiologic mechanisms that lead to the development of laminitis.     

Perceptions,practices, and consequences associated with foodborne pathogens and the feeding of raw meat to dogs

This study explored the impact of feeding raw meat to dogs on the fecal prevalence of several enteric bacterial zoonotic pathogens. Campylobacter jejuni was isolated from 1/42 (2.6%) raw meat-fed dogs. Salmonella enterica was isolated from 2/40 (5%) of the raw meat feeds, 6/42 (14%) raw meat-fed dog feces, none of the dogs that did not receive raw meat (P = 0.001), 4/38 (10.5%) of the vacuum cleaner waste samples from households where raw meat was fed, and 2/44 (4.5%) of vacuum cleaner waste samples from households where raw meat was not fed to dogs (P = 0.41). Responses to a questionnaire probing practices and beliefs regarding raw meat feeding that was administered to dog owners demonstrated that dog owners may either not be aware or refuse to acknowledge the risks associated with raw meat-feeding; thus, they may neglect to conduct adequate intervention strategies to prevent zoonoses among themselves and their families.     

Functional Activity of Frozen Thawed Chinchilla lanigera Spermatozoa Cryopreserved with Glycerol or Ethylene Glycol

The cryopreservation of spermatozoa constitutes a valuable tool for the captive breeding management of valuable and/or threatened species. Chinchilla lanigera is a species almost extinct in the wild, and the domestic counterpart has one of the most valuable pelts in the world. The objectives of this study were to: (i) compare the functional activity of post‐thawed chinchilla spermatozoa cryopreserved at ?196°C either with glycerol (G) or ethylene glycol (EG) as cryoprotectants (1 m final concentration) and (ii) investigate the effects of incubating the gametes for 4 h in the presence or in the absence of the cryoprotectants; evaluations were performed taking into account motility, viability, response to hypo‐osmotic shock and acrosome integrity of the cells. Parameters reflecting post‐thaw (0 h) sperm functional activity were significantly lower than those of freshly ejaculated gametes. When comparing the cryoprotectant efficiency of G vs EG, neither cryoprotectant agent offered appreciable advantages. After 4 h of incubation, in the presence or absence of the cryoprotectant agent, a rapid and significant decrease was found in all functional parameters and remained at ～ 20–30% motile, viable and viable acrosome intact cells. Viability was significantly lower when the cryoprotectant was removed from the media (possibly due to the centrifugation process). With respect to the maintenance of sperm membrane integrity, only ～ 10% of cells showed membrane resistance to hypo‐osmotic conditions after the 4 h incubation period. These results constitute new insights for cryopreservation protocols and the development of assisted reproductive techniques in this species.     

An allogeneic hybrid-cell fusion vaccine against canine mammary cancer

Mammary cancer is among the most prevalent of canine tumors frequently resulting in death due to metastatic disease. Most tumors fail to raise an effective immune reaction making improving immune recognition a priority. Hybrid-cell fusion strategies have been employed to load dendritic cell populations with tumor cell antigens to stimulate immune recognition; however, recovery, heterogeneity and quality of primary cells from patients present enormous challenges. We employed allogeneic cell lines to develop an improved hybrid-cell fusion strategy and evaluated immune reactions in normal laboratory beagles. Such a strategy relies on enhanced immune recognition of allogeneic tumor cell antigens by antigen presenting cells. Optimized PEG-promoted fusions between uniquely stained canine mammary tumor CMT12 or CMT28 cells and a dendritic cell-like DH82 cell fusion partner resulted in greater than 40% hybrid-cell fusion populations by flow cytometry and fluorescence microscopy. Hybrid-cell fusions were delivered by direct ultrasound guided injection into popliteal lymph nodes of laboratory beagles. Only hybrid-cell fusions provided statistically significant enhancement of cell-mediated immunity ((51)Cr-release assay) compared to innate reactions in na?ve vehicle injected dogs while dogs vaccinated with either single cell component alone did not. Vaccination with hybrid-cell fusions enhanced IFN-gamma expression in sorted CD8+ and CD4+ cells but not in CD4-/CD8- cells consistent with a CTL response. Cell-mediated immune assays revealed strong reactions against matched (vaccine component) CMT cells and unmatched CMT cells indicative of an immune response to mammary cancer antigens common to both cell lines. These results provide proof of principle for development of an allogeneic vaccination strategy against canine mammary cancer.     

Cartilage-derived retinoic acid-sensitive protein in equine synovial fluid from healthy and diseased joints

Reasons for performing study: More sensitive and specific diagnostic methods for early detection of changes in the joint cartilage are needed. Cartilage‐derived retinoic acid‐sensitive protein (CD‐RAP) is a potential marker of cartilage synthesis and regeneration. This is the first study on equine CD‐RAP. Objectives: To evaluate the ability of a commercially available human sandwich ELISA assay to detect equine CD‐RAP in synovial fluid from healthy and diseased joints. Methods: Synovial fluid was collected from 28 horses with no signs of joint disease and from 5 with induced inflammatory arthritis. CD‐RAP concentrations were measured using a human CD‐RAP ELISA. Intra‐ and interassay imprecision of the assay were evaluated by multiple measurements on pools of equine synovial fluid. Assay inaccuracy was determined by linearity under dilution. Results: The assay showed moderate to large intra‐ and interassay variation when applied to equine synovial fluid. Equine CD‐RAP was detected in synovial fluid from healthy horses ranged at 8.2–52 ng/ml. Repeated arthrocentesis (after injection of isotonic saline), age, joint or gender did not significantly affect CD‐RAP concentrations. Twelve hours after intra‐articular injection of lipopolysaccharide, concentrations of CD‐RAP were significantly lower than after injection of isotonic saline and remained significantly lower until the end of the study at 144 h. Conclusion and potential relevance: The assay is suitable for longitudinal monitoring of CD‐RAP concentration in individual horses. Disease significantly influenced CD‐RAP levels. Similar to previous results obtained in man, CD‐RAP seems to be a marker of cartilage synthesis and/or regeneration in horses.     

Improving the Reproductive Efficiency, Pregnancy Diagnosis and Monitoring the Resumption of Luteal Activity in Indigenous Damascus Goats

Seventy‐five female Damascus goats aged between 1.5 and 5.5 years were used to evaluate the effectiveness of the intravaginal sponges and prostaglandin analogue on oestrous synchronization and fecundity; to diagnose pregnancy and to monitor the resumption of the luteal activity. Females were divided randomly, during the breeding season, into three equal groups, S, P and C. Females in group S were fitted with sponges containing 45 mg of flugestone acetate (FGA) for 14 days and injected with pregnant mare serum gonadotrophin (PMSG) at the sponge withdrawal. Females in group P were given two injections of prostaglandin F2α analogue at 11‐day intervals, whereas females in group C (control) received no treatment. The results showed that there was a significant difference (p < 0.001) in oestrous exhibition between females in group S as compared with those in groups P and C, with means being 30 ± 10, 172 ± 115 and 217 ± 75 h for groups S, P and C, respectively. Kidding rates resulting from the first and all matings were 80 and 88, 52 and 88, and 68 and 80% for groups S, P and C, respectively. Fecundity rates were 215, 175 and 180% for groups S, P and C, respectively, with a significant difference (p < 0.05) between the S and both P and C groups. Using an ultrasound pregnancy detector performed on days 57 ± 3 after mating, positive pregnancy diagnosis was 93.3% and 100% for non‐pregnancy. Females in the control group showed functional corpus luteum starting in September. It is concluded that FGA sponges plus PMSG treatment could be successfully used to synchronize oestrus and improve fecundity; whereas prostaglandin treatment was not effective to synchronize oestrus. It is also concluded that pregnancy can be diagnosed accurately and successfully using an ultrasound pregnancy detector. In addition, ovarian activity in the Damascus goat in Syria resumes in September.