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.

Virulence-associated genes have been recognised and detected in Campylobacter species. The majority of them have been proven to be associated with pathogenicity. This study aimed to detect the presence of virulence genes associated with pathogenicity and responsible for invasion, expression of adherence, colonisation and production of the cytolethal distending toxin (cdt) in Campylobacter jejuni and Campylobacter coli. Commercial chicken faecal samples were randomly sampled from chicken farms within the Durban metropolitan area in South Africa. Furthermore, human clinical Campylobacter spp. isolates were randomly sampled from a private pathology laboratory in South Africa. Out of a total of 100 chicken faecal samples, 78% (n = 78) were positive for Campylobacter growth on modified charcoal cefoperazone deoxycholate and from the random laboratory collection of 100 human clinical isolates, 83% (n = 83) demonstrated positive Campylobacter spp. growth following culturing methods. These samples were screened for the presence of the following virulence genes: cadF, hipO, asp, ciaB, dnaJ, pldA, cdtA, cdtB and cdtC. As expected, the cadF gene was present in 100% of poultry (n = 78) and human clinical isolates (n = 83). Campylobacter jejuni was the main species detected in both poultry and human clinical isolates, whilst C. coli were detected at a significantly lower percentage (p < 0.05). Eight per cent of the C. jejuni from human clinical isolates had all virulence genes that were investigated. Only one C. coli isolate demonstrated the presence of all the virulence genes investigated; however, the pldA virulence gene was detected in 100% of the C. coli isolates in poultry and a high percentage (71%) in human clinical C. coli isolates as well. The detection of cdt genes was found at higher frequency in poultry than human clinical isolates. The high prevalence rates of virulence genes detected in poultry and human clinical isolates demonstrate their significance in the pathogenicity of Campylobacter species.

Medicinal herbal feed that used as feed additives are widely applied in livestock production and may be applicable to aquaculture production systems. The present study explores the modulatory effects of incorporation of herbal combination of black cumin seeds (Nigella sativa), peppermint (Mentha piperita) and fennel (Foeniculum vulgare), on African Sharptooth Catfish, Clarias gariepinus. For this purpose, a total of 72 catfish were randomly distributed into 6 groups of 12 fish each and fed on the experimental diets for 45 successive days. Fish of first group were fed on basic diet only while, those of the second one were fed on basic diet supplemented with 0.5% from each medicinal plant while, other 4 groups were subjected to challenge experiment by injection with Vibrio vulnificus. The third group was kept non-injected, fourth group was injected by saline, fifth and sixth groups were injected by Vibrio vulnificus after feeding with basic diet and basic diet with herbs, respectively. Results proved that, total antioxidant capacity, serum total proteins, albumins and globulins were observed to be significantly higher in the treated group as compared to the control. Lymphocytes percent increased significantly in herbal fed group comparing to the control group while, neutrophils percent decreased significantly. Although serum urea level was not affected by herbal supplement, serum creatinine level was decreased significantly. Body weight gain of C. gariepinus increased significantly after herbal administration. Catfish challenged with a Vibrio vulnificus isolate and received the herbal diet showed less mortality than the control group. Fish fed on the herbal diet exhibited normal histological structure of liver, kidney and spleen. In conclusion, based on the current results together with the low cost and the potential antioxidant and immune effects of mixed herbal medicinal plants used in the study, it is recommended to be used in fish feed to diminish the mortalities caused by some aquatic pathogens.

The experiment was designed to investigate the immunomodulating effect of lector 50 on general health and immune response of broiler chicks to Mycoplasma gallisepticum vaccination in commercial broiler chicks. the obtained results reveled significantly higher effects on body weight ,bursal, and thymic index on lector treated group of chickens, while no effects on spleen index. Also significant improvement in total and differential leukocytic count as well as significantly higher antibody titer was detected by ELISA in lector 50 treated groups.