Application of synthetic matrix metalloproteinases (MMPs) inhibitors, such as doxycycline is one of the possible therapeutic options for osteoarthritis. However, little is known about the protective mechanism of doxycycline in equine models on MMPs inhibitors as well as on serum amyloid A (SAA) gene expression. This study investigated the effects of doxycycline on mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13, and SAA of equine fibroblast-like synoviocytes (FLSs). The FLSs were established from synovial fluids of clinically normal metacarpophalangeal joints of 6 skeletally mature horses. The cells were treated with either 10 or 100 μg/mL of doxycycline for 48 h. The mRNA expression of MMP-1, MMP-2, MMP-9, MMP-13, and SAA were assessed using real-time polymerase chain reaction (PCR). Treatment with doxycycline resulted in significantly decreased mRNA expression of MMP-1 in FLSs at both concentrations (P = 0.001). No significant differences were detected among groups for MMP-2, MMP-9, and MMP-13 (P > 0.05). Only a tendency towards a decrease in mRNA expression level of SAA in the presence of doxycycline could be detected. Doxycycline inhibits MMP-1 gene expression at the transcript level. These findings indicate that doxycycline can protect the articular environment through inhibition of MMP-1 at transcript level.

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.

OBJECTIVE To compare tensile strength and time to completion of body wall closure among 3 suture patterns. SAMPLE Eighteen 5 × 5-cm leather specimens and sixty-eight 5 × 5-cm full-thickness tissue specimens from the ventral portion of the abdominal body wall of 17 canine cadavers. PROCEDURES During experiment 1 of a 2-experiment study, each leather specimen was cut in half and sutured with a simple interrupted or simple continuous pattern or continuous pattern with intermittent Aberdeen knots (intermittent Aberdeen pattern). During experiment 2, 4 tissue specimens were obtained from each cadaver; the linea alba of 3 specimens was incised and closed with 1 of the 3 suture patterns evaluated in experiment 1, and the fourth specimen was left intact as a control. All leather and tissue specimens underwent mechanical testing. Time to completion, mode of failure, and maximum force at failure (Fmax) were compared among the suture patterns. RESULTS In experiment 1, the mean Fmax for the simple continuous and intermittent Aberdeen patterns was significantly greater than that for the simple interrupted pattern. In experiment 2, the mean Fmax for specimens obtained cranial to the umbilicus was greater than that for specimens obtained caudal to the umbilicus, and the mean time to completion for both continuous suture patterns was significantly less than that for the simple interrupted pattern. Most (34/51) sutured tissue specimens failed because the suture cut through the tissue at the suture-tissue interface. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that the intermittent Aberdeen pattern may be an alternative for body wall closure in dogs.

OBJECTIVE To measure effects of oral Akkermansia muciniphila administration on systemic markers of gastrointestinal permeability and epithelial damage following antimicrobial administration in dogs. ANIMALS 8 healthy adult dogs. PROCEDURES Dogs were randomly assigned to receive either A muciniphila (109 cells/kg; n = 4) or vehicle (PBS solution; 4) for 6 days following metronidazole administration (12.5 mg/kg, PO, q 12 h for 7 d). After a 20-day washout period, the same dogs received the alternate treatment. After another washout period, experiments were repeated with amoxicillin-clavulanate (13.5 mg/kg, PO, q 12 h) instead of metronidazole. Fecal consistency was scored, a quantitative real-time PCR assay for A muciniphila in feces was performed, and plasma concentrations of cytokeratin-18, lipopolysaccharide, and glucagon-like peptides were measured by ELISA before (T0) and after (T1) antimicrobial administration and after administration of A muciniphila or vehicle (T2). RESULTS A muciniphila was detected in feces in 7 of 8 dogs after A muciniphila treatment at T2 (3/4 experiments) but not at T0 or T1. After metronidazole administration, mean change in plasma cytokeratin-18 concentration from T1 to T2 was significantly lower with vehicle than with A muciniphila treatment (−0.27 vs 2.4 ng/mL). Mean cytokeratin-18 concentration was lower at T1 than at T0 with amoxicillin-clavulanate. No other significant biomarker concentration changes were detected. Probiotic administration was not associated with changes in fecal scores. No adverse effects were attributed to A muciniphila treatment. CONCLUSIONS AND CLINICAL RELEVANCE Detection of A muciniphila in feces suggested successful gastrointestinal transit following oral supplementation in dogs. Plasma cytokeratin-18 alterations suggested an effect on gastrointestinal epithelium. Further study is needed to investigate effects in dogs with naturally occurring gastrointestinal disease.

A total number of 140 Egyptian breed donkeys were used in this study. Animals are classified into two groups according to the sex. Each group was further subdivided into four subgroups according to their ages. All animals were proved to be health by clinical and laboratory examinations. Two blood samples were collected from each donkey, one with anticoagulant and the other without anticoagulant for obtaining clear non-haemolysed serum. Various tests were conducted to measure the values of some blood contents. It was clear that total RBCs count, hemoglobin content and packed cells volume showed marked decrease with the increase of age. Significant difference in RBCs count between some groups and highly significant difference in Hb and PCV contents between another groups. Gradual elevation in the values of total leucocytes count from one month up to 10 years old was observed. Marked decrease in total WBCs count was reported in animals of both sexes on 10-20 years old. This denotes that Significant and highly significant differences appear in total WBCs count between animal groups. The biochemical parameters revealed highly significant difference in the total protein and albumin in some groups of male animals. Non significant fluctuation was observed in blood serum calcium, phosphorus and magnesium regarding the age and sex factors. In conclusion, it was clear that both age and sex factors has a marked influence on some blood contents in Egyptian donkeys.

This work was planned to investigate the bacteria isolated from broiler chickens head suffered from naked eye pathological lesions. Out of 200 examined head lesions, the result revealed that the major pathogens associated with swollen head syndrome (SHS) were Escherichia coli, Streptococcus dysgalactiae and Pseudomonas aeruginosa. Antimicrobial sensitivity pattern against 11 different antimicrobials proved that isolates were resistant to most of the tested antimicrobial agents. PCR was applied on 4 MDR E. coli, 4 S. dysgalactiae and 2 P. aeruginosa for detection of some resistance and virulence genes. The results of E. coli isolates revealed that blaTEM gene was the most prevalent in all isolates (100%) followed by tetA (A), aada1, aada2 and aacC genes. Meanwhile tetA (B) gene was found in 3 (75%), while aadB gene was not detected in any isolates. All S. dysgalactiae proved to harbour 16srRNAgene also all S. dysgalactiae were 100% positive for tuf gene followed by speF gene which found in 2 isolate (50%). The results of PCR of P. aeruginosa isolates revealed that toxA gene was the most prevalent gene found in all isolates (100%) followed by lasI. Then, phzM gene was found in one isolate (50%).

The current study was performed to evaluate the effect of acute and chronic hepatotoxicity induced by paracetamol and thioacetamide respectively on serum hormonal levels and biochemical parameters. Female Wistar albino rats were divided into 3 equal groups (C), (P) and (T). Group (C) were kept as control, group (P) were received paracetamol orally (500 mg/kg b.wt) daily for 15 days and those of group (T) were injected thioacetamide (200 mg/kg b.wt) intraperitonialy twice/ week for 90 days. In P group, results revealed significant elevation in liver enzyme activities (ALT, AST and ALP), T4, insulin (7th day), estrogen (7th and 15th days), triglycerides (7th day) and cholesterol levels throughout the experiment while serum proteins and T4 (15th day) showed significant decreased values. Whereas, at 90th days of chronic intoxicated group (T) resulted in significant elevation in liver enzyme activities (ALT, AST and ALP), bilirubin, estrogen, T4, triglycerides (60th and 90th days) and T3 (120th day). While the levels of T4 and cortisol (60th day), serum total protein, albumin, globulin (90th day) and insulin (120th day) showed significant decreased values when compared to control group. In conclusion, both paracetamol and thioacetamide cause different degrees of damage in liver of rats leading to clear changes in their hormonal and biochemical profiles.