Effects of temperature and storage time on canine bone-transfixation pin specimens were tested by comparing pin pull-out forces. A total of 16 femurs from 8 mature dogs were tested. Five nonthreaded Steinmann pins were placed through both cortices in the diaphysis of each femur. The femurs were then sectioned transversely between each pin, with a bonepin specimen placed evenly into each of 5 groups prior to biomechanical testing. Four bone-pin specimen groups were stored at -20 or -70 C for 14 or 28 days, while 1 specimen group was immediately tested. Pull-out forces for frozen groups were compared with pull-out forces for the fresh group. Using two-way ANOVA, there was no statistical difference in mean axial-extraction forces among bonepin specimen in any of the tested groups. It is concluded that acute pin pull-out forces are not significantly affected by freezing temperature or time. However, specimens stored at -20 C for as few as 14 days had a trend for increased pull-out forces, compared with freshly harvested specimens. Therefore, the authors recommend storage of bone-pin specimens at -70 C when possible.

Eight Holstein cows, 4 inoculated intracisternally in 1 quarter of the mammary gland with Escherichia coli and 4 noninfected controls, were administered ceftiofur sodium (3 mg/kg of body weight, IV, q 12 hours) for 24 hours, beginning at 14 hours after inoculation of infected cows. All challenge-exposed cows became infected, with mean +/-SEM peak log10 bacterial concentration in milk of 5.03 +/-0.69 colony-forming units/ml. The infection resulted in systemic signs (mean peak rectal temperature, 41.5 +/- 0.3 C; anorexia; signs of depression) and local inflammation (mean peak albumin concentration in milk, 7.89 +/- 1.71 mg/ml). Ceftiofur was detectable in milk from all challenge-exposed cows, compared with only 1 of 4 noninfected cows, and the mean period after inoculation that ceftiofur was detectable in milk was longer (P < 0.05) in infected (147.7 +/- 27.5 hours) than noninfected cows (1.3 +/- 1.3 hours). However, maximal ceftiofur concentration attained in milk for all cows was 0.28 micrograms/ml, and was 0.20 micrograms/ml or less for all but 2 milk samples collected for 10 days after challenge exposure. Mean serum concentration of ceftiofur peaked at 1.0 +/- 0.3 micrograms/ml and 0.7 +/- 0.1 micrograms/ml for infected and noninfected COWS, respectively. After each ceftiofur dose, mean peak and trough concentrations of ceftiofur in serum did not differ between groups; however, concentration of ceftiofur in serum was higher at 7 hours after each dose in noninfected cows, suggesting more rapid clearance of the drug in infected cows. Ceftiofur was not detected in serum (< 0.05 micrograms/ml) of any cow at or after 120 hours following inoculation of infected cows Storage of serum samples at -20 C for 3 weeks resulted in a 98.8% decrease in ceftiofur activity, compared with that in fresh serum samples. Eighty-seven percent of this loss occurred 30 minutes after mixing serum and ceftiofur; thus, about 13% of the original activity was lost in storage.

Introduction The addition of low-molecular-weight antioxidants during the freezing process improves post-thaw sperm quality. The high antioxidant potential of cryopreserved semen could have a positive effect on the motility, viability, and energy status of sperm cells and their ability to bind to the zona pellucida of oocytes. The aim of the study was to determine the effects of different concentrations and combinations of vitamins E and C in a semen extender on selected quality parameters of frozen-thawed canine spermatozoa. Material and Methods The experimental material was the semen of four mixed-breed dogs. Sperm viability (motility, plasma membrane integrity, and mitochondrial function) was examined at 0, 60, and 120 min in semen samples supplemented with the extender and in the controls. Results Combined supplementation with vitamins C + E at a concentration of 200 + 200 μM /1 × 10⁹ spermatozoa had the most profound effect on total sperm motility, linear motility, and the percentage of spermatozoa with intact plasma membrane and active mitochondria. Conclusion The synergistic activity of vitamins E and C had a more beneficial influence on the quality of frozen–thawed sperm than these non-enzymatic antioxidants applied separately.

The current study was designed to evaluate the influence of three different ground vegetable oil seeds on various physico-chemical, microbiological and sensory attributes of chicken sausages from spent broiler breeder hen stored under frozen (-18±1°C) condition. Chicken sausages were incorporated with ground poppy, sesame and peanut seeds each at 10 % level separately as three treatments and were analyzed for their keeping quality along with high fat control. The mean values of FFA, TBARS, tyrosine values, standard plate count, psychrophilic count and yeast and moulds counts were significantly (P&lt;0.05) influenced by formulation during frozen storage and values increased significantly (P&lt;0.05) throughout the storage period in all chicken sausages. Chicken sausages with ground sesame scored lowest standard plate counts (5.66 log CFU/g) than any others after 180 days of storage at 18±1°C. Coli forms, lacto bacillus and anaerobic counts could not be detected during entire storage period. Scores for all sensory attributes decreased significantly (P&lt;0.05) during storage in all treatments irrespective of formulation. Study proved that, the three ground vegetable oil seeds were effective by delaying the lipid oxidation, microbial growth and preserving sensory quality during frozen storage and among them ground sesame seed was more potent to be a better preservative.

OBJECTIVE To evaluate the effect of presurgical storage conditions on leakage pressures of enterotomy sites closed with unidirectional barbed suture material in fresh, chilled, and frozen-thawed cadaveric canine jejunal specimens. SAMPLE 36 grossly normal jejunal segments obtained from 4 dog cadavers. PROCEDURES 9 jejunal segments were harvested immediately from each euthanized dog and randomly assigned to be tested within 4 hours after collection (fresh segments), stored at 4°C for 24 hours before testing (chilled segments), or stored at −20°C for 7 days and thawed at 21°C for 6 hours before testing (frozen-thawed segments). For leakage pressure testing, a 3-cm-long antimesenteric enterotomy was performed and repaired with 3-0 unidirectional barbed suture material in a simple continuous pattern in each segment. Time to complete the enterotomy, initial leakage pressure, maximum intraluminal pressure, and leakage location were recorded for each segment. RESULTS Mean ± SD initial leakage pressure for fresh, chilled, and frozen-thawed segments was 52.8 ± 14.9 mm Hg, 51.8 ± 11.9 mm Hg, and 33.3 ± 7.7 mm Hg, respectively. Frozen-thawed segments had significantly lower mean initial leakage pressure, compared with findings for fresh or chilled segments. Time to complete the enterotomy, maximum intraluminal pressure, and leakage location did not differ among groups. CONCLUSIONS AND CLINICAL RELEVANCE Leak pressure testing of cadaveric jejunal segments that are fresh or chilled at 4°C for 24 hours is recommended for enterotomy studies involving barbed suture material in dogs. Freezing and thawing of cadaveric jejunal tissues prior to investigative use is not recommended because leak pressure data may be falsely low.

OBJECTIVE To compare effectiveness of glycerol, dimethyl sulfoxide (DMSO), and hydroxyethyl starch (HES) solutions for cryopreservation of avian RBCs. SAMPLE RBCs from 12 healthy Ameraucana hens (Gallus gallus domesticus). PROCEDURES RBCs were stored in 20% (wt/vol) glycerol, 10% (wt/vol) DMSO freezing medium, or various concentrations of HES solution (7.5%, 11.5%, and 20% [wt/vol]) and frozen for 2 months in liquid nitrogen. Cells were then thawed and evaluated by use of cell recovery and saline stability tests, cell staining (7-aminoactinomycin D and annexin V) and flow cytometry, and scanning electron microscopy. RESULTS Percentage of RBCs recovered was highest for 20% glycerol solution (mean ± SE, 99.71 ± 0.04%) and did not differ significantly from the value for 7.5% HES solution (99.57 ± 0.04%). Mean saline stability of RBCs was highest for 10% DMSO (96.11 ± 0.25%) and did not differ significantly from the value for 20% HES solution (95.74 ± 0.25%). Percentages of cells with 7-aminoactinomycin D staining but without annexin V staining (indicating necrosis or late apoptosis) were lowest for 10% DMSO freezing medium (3%) and 20% glycerol solution (1%) and highest for all HES concentrations (60% to 80%). Scanning electron microscopy revealed severe membrane changes in RBCs cryopreserved in 20% HES solution, compared with membrane appearance in freshly harvested RBCs and RBCs cryopreserved in 10% DMSO freezing medium. CONCLUSIONS AND CLINICAL RELEVANCE Cryopreservation of avian RBCs with HES solution, regardless of HES concentration, resulted in greater degrees of apoptosis and cell death than did cryopreservation with other media. Transfusion with RBCs cryopreserved in HES solution may result in posttransfusion hemolysis in birds.

The aim of this study was to compare the effects of cryopreservation at approximately -196 °C in liquid nitrogen (N) and freezing at approximately -20 °C in a freezer, on the viability and survival of eight different mastitogenic bacteria inoculated in milk. Bacteria were frozen at approximately -20 °C in a freezer and cryopreserved at approximately -196 °C in liquid nitrogen. An effective preservation method was needed for follow-up samples from cows identified in the South African National Milk Recording Scheme (NMRS) with somatic cell counts above 250 000 cells/mL milk. The organisation responsible for sample collection of the NMRS milk samples also provides producers with liquid nitrogen for their semen flasks at the collection sites. This existing mode of storage and transport could therefore be utilised. Ten samples of each organism were thawed and cultured bi-weekly until week 18 for both temperature treatments. An additional sampling was performed at week 30 for samples frozen at approximately -20 °C. Freezing and cryopreservation did not impair subsequent isolation of Streptococcus dysgalactiae, Streptococcus uberis, Enterococcus faecalis, Staphylococcus aureus (STH) (phage type lytic group III) or Sta. aureus (STA) (phage typed, other than lytic group III). Survival was indicated by the isolation of bacteria from samples, and viability by the strength of growth of the bacteria isolated. The survival of Streptococcus agalactiae decreased after week 12 and Escherichia coli after week 16 of freezing, but both organisms survived under cryogenic preservation until week 18. Coagulase-negative staphylococci survived until week 18 for both freezing and cryogenic preservation. Both storage methods could thus contribute to the improvement of a pro-active approach towards udder health management in South African dairy herds.

An ELISA containing lipoarabinomannan (LAM) antigen was used to detect antibodies in milk and serum for diagnosis of Mycobacterium paratuberculosis infection in dairy cattle. In experiment 1, milk and serum samples were obtained from 25 cows, and subjected to LAM ELISA testing immediately, and after 1 year of storage at -70 C. Milk samples, with and without a commonly used chemical preservative, were tested. There was no significant difference in LAM ELISA results between fresh and frozen samples or between preserved and unpreserved milk samples. In experiment 2, milk samples were collected daily from 30 cows over a 14-day period. The day-to-day coefficient of variation was 0.19 for milk LAM ELISA and was 0.15 for serum LAM ELISA, with no statistically significant time effect detected. In experiment 3, single milk, serum, and fecal samples were obtained from 764 cows. The fecal samples were cultured for M paratuberculosis to identify infected cows, and the serum and milk samples were subjected to LAM ELISA testing. Results were compared, using the area under the receiver operating characteristic curves. The milk LAM ELISA had specificity (+/- 95% confidence limits) of 87 +/- 8.1% when the cutoff was set at 50% sensitivity, and specificity of 83 +/- 9.1% when sensitivity was set at 60%. The area under the receiver operating characteristic curve was 0.85 +/- 0.03 for the milk ELISA and 0.75 +/- 0.02 for the serum ELISA. In this population of cattle, the milk LAM ELISA had comparable accuracy to serum LAM ELISA, although the milk LAM ELISA was slightly less reproducible (higher coefficient of variation).