A total of 1317 samples were collected; 1164 serum samples, 122 milk samples, 24 lymph nodes and 7 aborted foeti from buffaloes in 10 Governorates from farms and villages in Upper Egypt. The serological tests used for the diagnosis of brucellosis on blood sera were the Rose Bengal plate (RBT) , Buffered acidified plate antigentest (BABAT), EDTA modifiedstandard tube agglutination test (MSAT), Revanol test (RT). On the other hand, the milk ring test (MRT) was performed on buffalo-cow's milk. Suspected colonies were stained with Gram, s stain and Modified ZeilNeelson stain. The isolated Brucella organisms on antibiotic free Brucella agar medium were subjected to the following tests for biochemical identification tests as CO2requirement, H2S production, Urease activity, growth in the presence of dyes,The indirect solid phase ELISA technique was carried out according to serum and milk samples.Agar gel immune diffusion test (AGID) and PCR applied on isolated Brucella strains. The results of the serological tests wereRose Bengal test 34.7%, BAPA (37%), Revanol test (28.2%),modified SAT (23.7%), indirect ELISAwere (32.3%) and AGPT (33.8%)in this study.Brucellaorganisms from lymph nodes of slaughtered buffaloes by culturing method showed that 3 (13.64%) isolates(2) of B. melitensisbiovar 3 and (1)B. abortusbiovar1. The isolated strain from aborted foeti was one isolate (14.29%) typed as B.melitensisbiovar 3. isolated only from Beni-Suef.By milk ring test (MRT) milk samples were 10 (8.20%) of B. melitensis biotype 3. A multiplex was format that will allow the rapid identification of Brucella spp., B. abortus, and B.melitensis in a single test within 2 to 3 h. B. melitensis was identified at 731bp and B. abortus identified at 498bp. Finally, we made measures of the control program for eradication of brucellosis in buffaloes by a reasonable system of compensation, Veterinarians for field work and state laboratories capable of serological techniques.Also, information technology solutions and further logistic means as animal identification techniques are in any governorates in Egypt.

Presently neither the specification of rumen cannula for small ruminants is reported nor is it commercially available in Egyptian market. Therefore, fabrication of ruminal cannula for sheep and surgical procedure for its implantation are described in this paper. The device was adapted to allow sampling of entire ruminal contents via cannulas with different diameters, which tightly sealed within ruminal fistula to ensure cleaner, achieve easier nursing of operated animals, and maintain more normal ruminal environment. The ruminal cannula was applied into the sheep by one-stage operation. It has been successfully used in 11 ram (3-5 year-old) for 16 months without problems and caused no complications.

Fungal diseases of poultry have become problematic as bacterial and viral diseases. This study was designed to investigate the prevalence of fungal agents in broiler chickens and their environment. The prevalence of fungal isolation from broiler chickens was 21.6% including 12.8% moulds and 8.8% yeast while the prevalence of fungal isolation from the environment was 46.8% including 25.5% moulds and 21.3% yeast. Aspergillus species was the most prevalent moulds while C. albicans was the most prevalent yeast recovered from broiler chickens and their environment.

OBJECTIVE To assess insulin, glucagon, and somatostatin expression within pancreatic islets of horses with and without insulin resistance. ANIMALS 10 insulin-resistant horses and 13 insulin-sensitive horses. PROCEDURES For each horse, food was withheld for at least 10 hours before a blood sample was collected for determination of serum insulin concentration. Horses with a serum insulin concentration < 20 μU/mL were assigned to the insulin-sensitive group, whereas horses with a serum insulin concentration > 20 μU/mL underwent a frequently sampled IV glucose tolerance test to determine sensitivity to insulin by minimal model analysis. Horses with a sensitivity to insulin < 1.0 × 10(−4) L•min−1•mU−1 were assigned to the insulin-resistant group. All horses were euthanized with a barbiturate overdose, and pancreatic specimens were harvested and immunohistochemically stained for determination of insulin, glucagon, and somatostatin expression in pancreatic islets. Islet hormone expression was compared between insulin-resistant and insulin-sensitive horses. RESULTS Cells expressing insulin, glucagon, and somatostatin made up approximately 62%, 12%, and 7%, respectively, of pancreatic islet cells in insulin-resistant horses and 64%, 18%, and 9%, respectively, of pancreatic islet cells in insulin-sensitive horses. Expression of insulin and somatostatin did not differ between insulin-resistant and insulin-sensitive horses, but the median percentage of glucagon-expressing cells in the islets of insulin-resistant horses was significantly less than that in insulin-sensitive horses. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that, in insulin-resistant horses, insulin secretion was not increased but glucagon production might be downregulated as a compensatory response to hyperinsulinemia.

OBJECTIVE To evaluate effects of various concentrations of collagenase and dimethyl sulfoxide (DMSO) on yield of equine adipose-derived multipotent stromal cells (ASCs) before and after cryopreservation. SAMPLE Supragluteal subcutaneous adipose tissue from 7 Thoroughbreds. PROCEDURES Tissues were incubated with digests containing 0.1%, 0.05%, or 0.025% type I collagenase. Part of each resulting stromal vascular fraction was cryopreserved in 80% fetal bovine serum (FBS), 10% DMSO, and 10% Dulbecco modified Eagle medium F-12 and in 95% FBS and 5% DMSO. Half of each fresh and cryopreserved heterogeneous cell population was not immunophenotyped (unsorted) or was immunophenotyped for CD44+, CD105+, and major histocompatability complex class II (MHCII; CD44+-CD105+-MHCII+ cells and CD44+-CD105+-MHCII− cells). Cell proliferation (cell viability assay), plasticity (CFU frequency), and lineage-specific target gene and oncogene expression (reverse transcriptase PCR assays) were determined in passage 1 cells before and after culture in induction media. RESULTS Digestion with 0.1% collagenase yielded the highest number of nucleated cells. Cell surface marker expression and proliferation rate were not affected by collagenase concentration. Cryopreservation reduced cell expansion rate and CD44+-CD105+-MHCII− CFUs; it also reduced osteogenic plasticity of unsorted cells. However, effects appeared to be unrelated to DMSO concentrations. There were also variable effects on primordial gene expression among cell isolates. CONCLUSIONS AND CLINICAL RELEVANCE Results supported the use of 0.1% collagenase in an adipose tissue digest and 5% DMSO in cryopreservation medium for isolation and cryopreservation, respectively, of equine ASCs. These results may be used as guidelines for standardization of isolation and cryopreservation procedures for equine ASCs.

The objectives of the study were to: i) determine baseline microvascular perfusion indices (MPI) and assess their repeatability in healthy horses under general anesthesia, and ii) compare the MPIs of 3 microvascular beds (oral mucosa, colonic serosa, and rectal mucosa). Healthy adult horses were anesthetized and sidestream dark field microscopy was used to collect video loops of the oral mucosa, rectal mucosa, and colonic serosa under normotensive conditions without cardiovascular support drugs; videos were later analyzed to produce MPIs. Baseline MPI values were determined for each site, which included the total vessel density (TVD), perfused vessel density (PVD), portion perfused vessels (PPV), and microcirculatory flow index (MFI). Differences in MPIs between microvascular beds were not statistically significant. Repeatability of the measurements varied for each MPI. In particular, the site of sampling had a profound effect on the repeatability of the PPV measurements and should be considered in future studies.

OBJECTIVE To compare time to achieve vascular access (TTVA) between an ultrasound-guided technique (UST) and landmark-based technique (LMT) for central venous catheter (CVC) placement in healthy anesthetized dogs. ANIMALS 39 purpose-bred hounds. PROCEDURES Anesthetized dogs that were hemodynamically stable following completion of a terminal surgical exercise were enrolled in the study during 2 phases, with a 45-day intermission between phases. For each dog, a UST and LMT were used for CVC placement via each external jugular vein by 2 operators (criticalist and resident). The TTVA and number of venipuncture attempts and catheter redirections were recorded for each catheterization. Placement of the CVC was confirmed by contrast fluoroscopy. After euthanasia, a gross dissection was performed during which a hematoma score was assigned to the catheter insertion site. For each phase, nonlinear least squares estimation was used for learning curve analysis of the UST. RESULTS Median TTVA, number of venipuncture attempts and catheter redirections, and hematoma score did not differ significantly between the 2 operators for either technique. Median TTVA for the UST (45 seconds) was significantly longer than that for the LMT (7 seconds). Learning curve analysis indicated that 8 and 7 UST catheterizations were required to achieve performance stability in phases 1 and 2, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that the UST was comparable to the LMT for CVC placement in healthy dogs. The extra time required to perform the UST was not clinically relevant. Additional studies evaluating the UST for CVC placement in clinically ill dogs are warranted.

OBJECTIVE To retrospectively evaluate the epidemiological and morphological features and outcome of surgical treatment of incomplete ossification of the dorsal neural arch of the atlas (IODA) in dogs with atlantoaxial instability (AAI). ANIMALS 106 AAI-affected dogs that underwent ventral fixation of the atlantoaxial joint. PROCEDURES Medical records and CT images for each dog were reviewed. Dogs were allocated to 1 of 2 groups on the basis of the presence or absence of IODA or of dens abnormalities (DAs) in CT images. RESULTS Of the 106 dogs with AAI, 75 had and 31 did not have IODA; 70 had and 36 did not have DAs. Incomplete ossification was present in the cranialmost, central, or caudalmost portion of the dorsal neural arch of the atlas in 59, 39, and 28 dogs, respectively; 2 or 3 portions were affected in 29 and 11 dogs, respectively. The mean CT value (in Hounsfield units) for the midline of the dorsal neural arch of the atlas in dogs with IODA was significantly lower than that for the same site in the dogs without IODA. The mean age at surgery for dogs with central IODA was significantly higher than that of the non-IODA group. The severity of spinal cord injury before or after atlantoaxial ventral fixation did not differ between the IODA and non-IODA groups. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that concomitant DAs or IODA is common in dogs with AAI. In dogs with incomplete ossification in the central part of the dorsal neural arch of the atlas, surgical treatment of AAI generally occurs at a middle to advanced age.

OBJECTIVE To determine the in vitro effects of calcitriol on indicators of immune system function in blood samples collected from healthy dogs. SAMPLE Blood samples from 8 healthy adult dogs. PROCEDURES Blood samples were incubated with calcitriol (10(−7)M) or control substance for 24 hours. Afterward, lipopolysaccharide (LPS)-, lipoteichoic acid (LTA)-, and N-acetylmuramyl-l-alanyl-d-isoglutamine hydrate (MDP)-stimulated leukocyte production of tumor necrosis factor (TNF) and interleukin-10 (IL10) were measured with a canine-specific multiplex assay. Phagocytosis of opsonized Escherichia coli and leukocyte expression of constitutive toll-like receptor 4 (TLR4) were evaluated via flow cytometry. Blood samples from 3 dogs were used to create a concentration-response curve to evaluate whether the observed cytokine modulation was concentration dependent. RESULTS Incubation of canine blood samples with calcitriol resulted in significant decreases in LPS-, LTA-, and MDP-stimulated leukocyte production of TNF but not IL10. Blunting of TNF production was concentration dependent. Leukocyte calcitriol exposure had no significant effect on phagocytosis and TLR4 expression. CONCLUSIONS AND CLINICAL RELEVANCE These data indicated that calcitriol induced an anti-inflammatory shift in canine leukocytes exposed to LPS, LTA, and MDP in vitro, without altering phagocytosis or TLR4 expression. Thus, calcitriol could represent a novel candidate immunomodulatory treatment for dogs.