Comparison between commercial media and TRIS-egg yolk extender in the refrigerated storage of collared peccary semen at 17°C.

To assist in the conservation of collared peccary, it is important to strengthen semen processing protocols. The aim of this study was to compare the effects of different commercial extenders (BTS; NUTRIXcell+ and PRIMXcell Ultra) and TRIS + egg yolk on the functional and morphological aspects of collared peccary semen stored at 17 °C for 48 hours. Ten ejaculates obtained by electroejaculation were divided into 4 aliquots and diluted in the respective extenders, then stored in a biological incubator at 17 °C for 12, 24, 36, and 48 hours. The samples were evaluated for kinetic parameters, membrane functionality, membrane integrity, mitochondrial activity, morphology, and sperm-binding capacity. At the end of storage (48 h), promising results were found for motility parameters, with TRIS + egg yolk (71.0 ± 4.6%) being more efficient than NUTRIXcell+ (38.9 ± 10.9%) (P < 0.05) and similar to BTS (42.9 ± 11.9%) and PRIMXcell Ultra (46.8 ± 10.8%). The results for membrane integrity and mitochondrial activity were around ∼30-50%, with TRIS being the only extender to preserve both parameters (58.9 ± 5.3 and 59.2 ± 5.6%) for up to 48 hours, respectively (P < 0.05). Finally, the extenders could guarantee 60% membrane functionality and ∼ 60-70% normal sperm morphology, as well as similar binding capacity among the groups. In conclusion, TRIS + egg yolk is effective in preserving the sperm parameters of collared peccary semen at 17 °C for 48 hours, while PRIMXcell Ultra and BTS are viable alternatives for this purpose.

One-week storage of refrigerated bovine milk does not affect the size, concentration, or molecular properties of extracellular vesicles.

Milk extracellular vesicles (EV) have gained extensive attention as promising diagnostic and therapeutic tools. Pre-analytical raw milk storage at low temperatures is an ordinary and usually necessary step after sample collection. It is known that direct freezing of unprocessed whole milk contaminates the native pool of milk EV with other cell structures. However, less evidence is available regarding prolonged cooling at 4°C. The current study assessed whether pre-analytical storage of bovine raw milk for several days affected EV isolation and further analysis. To confirm the independence from the health status of the mammary gland, we analyzed milk samples stored at 4°C for 1, 2, 3, and 7 d past collection, respectively, from 2 quarters of the same cow with different somatic cell counts (SCC). Seven days of refrigeration did not change the milk EV size, concentration, or morphology. We did not detect any changes in the EV cargo regarding the amount of protein and RNA, nor in the specific EV markers TSG101, CD9, and CD81 in milk from quarters with high and low SCC. Overall, we observed fewer CD81 and CD9 markers in quarters with high SCC. Moreover, we found no reduction in the mastitis-related miRNA bta-miR-223-3p, suggesting that refrigeration for several days up to 1 wk is a possible storage option compatible with further EV analyses. The findings of this study enhance the confidence that milk EV are highly stable in the raw milk matrix.

Effects of time and temperature of storage on chemical and nutritional characteristics of raw milk for Provolone Valpadana PDO cheesemaking: a multivariate approach.

We evaluated the possibility of increasing the storage temperature of raw milk for Provolone Valpadana cheesemaking, to identify the most suitable conditions of time and temperature for a pre-maturation process. We used Principal Component Analysis (PCA) to analyze the overall effects of different storage conditions on chemical, nutritional and technological characteristics of the raw milk. Four different thermal storage cycles, two at fixed temperature/time (6 and 12°C for 60 h) and two with two-phase thermal cycle (10 and 12°C for 15 h, followed by refrigeration at 4°C for 45 h) were studied. Although a moderate heterogeneity among raw milks from the 11 producers of Provolone Valpadana cheese was observed, PCA revealed the critical aspects of the extreme storage conditions (60 h of refrigeration). Some samples resulted in anomalous behaviors, probably related to unexpected fermentation phenomena occurring with increasing storage temperature. The acidification and the increase in the contents of lactic acid, soluble calcium, and degree of retinol isomerization observed in the anomalous samples can compromise the technological functionality of milk. Conversely, the storage with a two-phase thermal cycle did not lead to variations in any measured characteristic, suggesting that mild refrigeration conditions (10 or 12°C for 15 h followed by 4°C for 45 h) could be a good compromise in favoring milk pre-maturation without altering its quality characteristics.

Effects of handling and storage on potassium concentration in plasma and serum samples obtained from cats.

OBJECTIVE: To compare potassium concentrations in feline plasma and serum samples analyzed promptly after collection or after 20 to 28 hours of refrigerated storage. ANIMALS: 41 cats. PROCEDURES: A venous blood sample was obtained from each cat. Aliquots were placed in 2 tubes without anticoagulant (blood was allowed to clot to derive serum) and 2 tubes with heparin (to derive plasma). One serum and 1 plasma sample were kept at room temperature and analyzed within 60 minutes after collection (baseline); the other serum and plasma samples were analyzed after 20 to 28 hours of refrigerated storage. At both time points, serum and plasma potassium concentrations were measured. RESULTS: Median baseline serum potassium concentration (4.3 mmol/L) was significantly higher than median baseline plasma potassium concentration (4.1 mmol/L). The median difference between those values was 0.4 mmol/L (95% CI, 0.2 to 0.5 mmol/L). Compared with their respective baseline measurements, the median serum plasma concentration (4.8 mmol/L) and median plasma potassium concentration (4.6 mmol/L) were higher after 20 to 28 hours of refrigeration. CLINICAL RELEVANCE: Results indicated that with regard to potassium concentration in feline blood samples, clotting or refrigerated storage for 20 to 28 hours results in a significant artifactual increase. Detection of an unexpectedly high potassium concentration in a cat may represent pseudohyperkalemia, especially if the blood sample was placed in a no-additive tube, was stored for 20 to 28 hours prior to analysis, or both.

Osmotic tolerance of rabbit spermatozoa is affected by extender composition and temperature.

Sperm osmotic adaptability to anisosmotic conditions is important for sperm epididymal maturation, motility activation at ejaculation, and female tract colonization, or for conducting technological procedures such as cryopreservation. Several factors affect this adaptability, including the fluid composition that contributes to water flow dynamics, and the temperature at which osmotic stress is initiated. This study was designed to investigate the effect of medium composition (electrolyte- or sugar-based extender) and temperature (25 and 5 °C) on rabbit sperm adaptability to anisosmotic conditions. Rabbit spermatozoa, therefore, were diluted at both temperatures (25 and 5 °C) in electrolyte- or sugar-based media at increasing osmotic conditions (100 to 1,000 mOsm/kg), and values for sperm variables (sperm kinetics, membrane integrity, mitochondrial membrane potential) were estimated as endpoints. Sperm kinetics seemed to be more sensitive to osmotic stress than membrane integrity or mitochondrial function. The effect of moderate hypoosmotic stress did not differ when there was use of sugar- and electrolyte-based extenders at 25 °C (P > 0.05). In hyper-tonic conditions at 25 °C, the sugar-based extender was more effective in protecting sperm membrane integrity and mitochondrial function (P < 0.05). The lesser temperature made the differences more relevant because of the detrimental effect of hyperosmotic stress was more evident in the electrolyte-based extender at 5 °C (P < 0.05). The results from this study indicated rabbit spermatozoa have different adaptability to anisosmotic conditions induced by sugar- and electrolyte-based media and that the temperature at which the osmotic stress is initiated affects the cellular response.

Screening of frozen-thawed conditions for keeping nutritive compositions and physicochemical characteristics of goat milk.

Frozen milk can help producers overcome the seasonality of goat milk production, low goat production and short lactation periods, and avoid discarding milk during some special periods. We investigated effects of combination between freezing (cryogenic refrigerator of -16 to -20°C or ultra-cryogenic refrigerator of -76 to -80°C) and thawing (homeothermy of 20 to 25°C or refrigeration of 2 to 4°C) on nutritive compositions and physicochemical characteristics of raw goat milk during storage period (80 d). Compared with fresh goat milk, the frozen-thawed milk decreased contents of fat, protein, and lactose, as well as surface tension and stability coefficient, whereas increased effective diameter and polydispersity index. The average values of color values (L*, a*, and b*) in 4 group samples changed from 83.01 to 82.25, -1.40 to -1.54, 3.51 to 3.81, respectively, and the ΔE of most samples did not exceed 2. In contrast to the other 3 frozen-thawed treatments, goat milk treated with ultra-cryogenic freezing-homeothermic thawing (UFHT) possessed higher fat (5.20 g/100 g), smaller effective particle diameter (0.32 µm), and the lowest polydispersity index value (0.26). The color and confocal laser scanning microscopy images of UFHT were similar to those of fresh goat milk, illustrating UFHT was the optimal approach to maintain the natural quality of goat milk. Our finding provides a theoretical basis for producers to freeze surplus milk.

An alternative cold chain for storing and transporting East Coast fever vaccine.

East Coast fever (ECF) is an often fatal, economically important cattle disease that predominantly affects eastern, central, and southern Africa. ECF is controlled through vaccination by means of simultaneous injection of oxytetracycline and cryogenically preserved stabilate containing live, disease-causing parasites. Storage and transportation of the stabilate requires liquid nitrogen, a commodity that is commonly unreliable in low-resource settings. Here we show that storage of conventionally prepared stabilate at -80 °C for up to 30 days does not significantly affect its ability to infect cultured peripheral blood mononucleated cells or live cattle, suggesting an alternative cold chain that maintains these temperatures could be used to effectively manage ECF.

Conservation of the internal quality of eggs using a biodegradable coating.

This study aimed to evaluate the effect of a pectin biofilm on the preservation of refrigerated and unrefrigerated eggs during 5 wk of storage based on egg weight loss, albumen height, Haugh unit (HU), and the yolk index (YI). A total of 1,200 nonfertile eggs from GLK Bankiva laying hens (40 wk of age), which were freshly laid and came from a single collection, were obtained from a model poultry rearing system (Planaltina, Federal District, Brazil) that meets all animal welfare criteria. The experimental outline was entirely randomized, with 20 treatments in a factorial scheme of 2 × 2 × 5, with 2 biofilm treatments (with and without) × 2 storage temperatures (refrigeration: 5°C and ambient: 25°C) × 5 storage periods (7, 14, 21, 28, and 35 d), with 12 repetitions per treatment. Starting from the third storage week, increased weight loss (%) was observed in noncoated eggs (4.46 ± 1.06; 5.61 ± 1.37; 6.93 ± 1.66%) compared with biofilm-coated eggs (3.57 ± 1.26; 4.74 ± 1.8; 6.05 ± 2.21%), respectively. The HU variation in the pectin-coated eggs (86.84-78.02) was smaller than that in the noncoated eggs (83.01-64.36) between the beginning (7 d) and the end (35 d) of the experimental period. Eggs with and without biofilm stored in the refrigerator presented average HU values of 91.26 ± 6.27 and 88.35 ± 6.96, respectively. In contrast, when kept at room temperature, eggs with the coating presented higher HU values (71.27 ± 10.78) than eggs without the coating (59.11 ± 15.97). Coated eggs (0.37 ± 0.16) showed higher YI values than noncoated eggs (0.35 ± 0.16). A pectin-based biofilm effectively maintained egg quality during the 35 d of storage.

Investigation of the effects of storage with preservatives at room temperature or refrigeration without preservatives on urinalysis results for samples from healthy dogs.

OBJECTIVE: To compare urinalysis results for canine urine samples stored in preservative-containing tubes at room temperature (20°C to 25°C [68°F to 77°F]) or refrigerated at 4°C (39.2°F) in plain glass tubes with results for the same samples immediately after collection. SAMPLES: Urine samples from 20 healthy dogs. PROCEDURES: Urine samples (1/dog) were divided into 6 aliquots (3 in preservative-containing tubes and 3 in plain glass tubes). Preservative-containing tubes were stored at room temperature and plain glass tubes were refrigerated. Urinalysis was performed 0, 24, and 72 hours after collection. Results for both storage conditions were compared with results for a reference sample (the 0-hour [immediate post-collection] aliquot in a plain glass tube) by Spearman correlation analysis with pairwise tests for selected variables. RESULTS: Physical variables (urine color and turbidity with and without centrifugation) for both storage conditions had high (rs = 0.7 to 0.9) or very high (rs = 0.9 to 1.0) degrees of positive correlation with reference sample results at all time points, except for color at 24 hours. Similar results were found for all biochemical variables with storage up to 72 hours. For microscopic characteristics, correlation with reference sample results ranged from low or nonsignificant to very high under both storage conditions. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that if a delay in urinalysis is expected, use of the preservative-containing tubes evaluated in this study may be a viable option for sample storage. Further research is warranted to assess direct comparability of results to those of freshly collected samples and use of these tubes to store samples from dogs with conditions affecting the urinary tract.

Molecular evidence of bovine herpesvirus 1 and 5 in cattle with suspected rabies in Rio Grande do Sul state, Brazil.

Rabies and herpetic encephalitis are the main viral infections in bovines with neurological symptoms. Bovine rabies has a high prevalence in Central and South America, while bovine encephalitis associated with herpesvirus is especially important in South America. Viral isolation is the classical way to confirm herpesvirus infection, but molecular evidence of the presence of the virus in affected animals is gaining importance in the diagnosis of the disease in the laboratory. This study investigated the presence of herpesvirus type 1 and 5 (BoHV-1 and BoHV-5) in 182 encephalon of rabies-suspected cattle in Rio Grande do Sul state (RS), Brazil using multiplex real-time polymerase chain reaction (mRT-PCR). The rabies virus was investigated by direct fluorescent antibody assay and intracerebral suckling mouse inoculation. The genomes of BoHV-1 and BoHV-5 were detected in 17% of samples. BoHV-5 and BoHV-1 were detected in 100% and 19% of BoHV positive samples, respectively, indicating the circulation of the pathogens in cattle herds in RS. The high Ct values and the absence of isolation suggest viral latency. Coinfection of herpesvirus and the rabies virus was detected in 28% of samples, although no significant association between pathogens was observed. Rabies was detected in 57.7% of suspected samples, confirming the importance of the disease in the state. Concerning the method by which samples were conserved, no significant difference was observed between the number of positive results in frozen and refrigerated samples.

Effects of gametes post-activation, egg storage and sperm-egg ratio on in vitro fertilization outcomes in the brown-marbled grouper (Epinephelus fuscoguttatus).

Due to some biological characteristics of the brown-marbled grouper, there are challenges in aquaculture enterprises focused on production of this fish. With the aim of facilitating captive breeding in brown-marbled grouper, effects on gamete post-activation, post-stripping egg storage and sperm to egg ratio on fertilization success were investigated. Results indicated eggs were fertile for 4 min after placement in seawater, thereafter, there was a reduction in fertilization. Initial curvilinear velocity (VCL) and total motility of fresh sperm was 129.3 ±â€¯3.7 µm/s and 92.9 ±â€¯2.4 %, both of which decreased as time elapsed post-activation, however, there was no significant difference in fertilization until 6 min post-activation of sperm. Compared with fertilization and hatching rates in eggs used immediately after collection (95.0 ±â€¯2.8 % and 90.1 ±â€¯4.2 %), in vitro storage of eggs had no detectable effect on fertilized egg and hatching rates until 60 min post-stripping (92.0 ±â€¯2.8 % and 89.9 ±â€¯2.7 %), indicating that immediate use of brown-marbled grouper eggs for fertilization is not necessary in aquaculture hatcheries. With regard to optimizing the sperm to egg ratio, there was acceptable fertilization with ratios ranging from 103 to 107, whereas there was a marked decrease in fertilization rate when there were lesser sperm concentrations. When the combined results from this study are considered, there could be improvements of artificial fertilization efficiency in the brown-marbled grouper with utilization of these findings in aquaculture enterprises focused on production of this fish.

Colloid centrifugation reduces bacterial load in chilled dog semen.

Conventional semen extenders contain antibiotics to prevent bacterial growth. Finding alternatives would be beneficial to minimize the development of bacterial resistance mechanisms. The aim of this study was to determine the effect of Single Layer Centrifugation (SLC) with Canicoll of dog semen on microbial load and sperm quality during cooled storage. Twenty-four ejaculates were obtained from healthy dogs by digital manipulation. Samples were diluted in Tris-citrate-fructose extender without antibiotics and divided into two treatment groups: SLC-selected samples and unselected samples. Sperm motility (CASA), viability and acrosome integrity (PI/FITC-PNA) as well as bacterial load of each microorganism species (colony-forming units/mL) were assessed at 0 and 48 h of storage at 4 °C. Results indicate SLC-selected dog spermatozoa have greater percentages of motility, viability and acrosome integrity (P < 0.05). Bacterial growth in SLC sperm samples was less (P < 0.05) than unselected samples. Removal of individual bacterial species varied from 91 % to 98 % for Escherichia coli (91.62 %), Streptococcus spp. (98.18 %), Staphylococcus spp.(95.33 %) and Pseudomonas spp. (92.50 %). In conclusion, the use of SLC with Canicoll has the potential to decrease bacterial load in chilled dog semen.

Detection of the mecA gene in Staphylococcus spp. isolated from the nasal cavity of pigs - Preliminary data / [Detecção do gene mecA em Staphylococcus spp. isolados da cavidade nasal de suínos - dados preliminares]

O objetivo deste estudo foi realizar a caracterização genética de Staphylococcus spp. resistentes a meticilina isolados de suínos. Foram coletadas 30 amostras de swab nasal de suínos, abatidos em um frigorífico com Serviço de Inspeção Federal. Os isolados foram submetidos a análises macro e microscópicas que, em seguida, para detectar a resistência bacteriana, foram submetidos a ensaios fenotípicos da sensibilidade aos antimicrobianos. Posteriormente, as amostras resistentes a oxacilina, foram submetidas à reação em cadeia pela polimerase (PCR) para verificar a presença do gene mecA. Das 30 amostras analisadas, foram isolados 12 (40%) Staphylococcus spp. coagulase positiva, e 18 (60%) coagulase negativa, e, dentre os isolados, 26 (86,66%) foram resistentes a oxacilina sendo possível detectar o gene mecA em seis (23%) amostras. Este estudo evidencia a presença de genes de resistência em microrganismos comuns a microbiota de animais de produção que podem ser transmitidos ao homem. Além de chamar a atenção para a frequência e quantidade de antimicrobianos aos quais estes animais são expostos durante toda sua vida, podendo ser considerado um problema para a saúde única.(AU)

Technical note: Effect of delayed analysis of cooled lithium-heparinized whole blood on the stability of ionized calcium, ionized magnesium, sodium, potassium, chloride, glucose, and lactate in samples from dairy cows.

The objectives of this study were to describe the stability of bovine whole-blood electrolytes, glucose, and lactate in samples collected in lithium heparin tubes and stored in thermoconductive modules immersed in ice water. A total of 99 Jersey cows (40 first-parity, 18 second-parity, and 41 third-parity or greater cows) from a commercial dairy farm in West Texas were enrolled between June and July 2018. Blood was collected from the jugular vein using a 60-mL polypropylene syringe and equally distributed into 5 spray-dried evacuated lithium heparin tubes. Baseline samples were analyzed within 90 s of collection using a benchtop blood gas analyzer. The remaining 4 tubes were stored in a thermoconductive, passive-temperature-regulating module inside a cooler with ice water. At 30 min and 2, 4, and 8 h post-collection, samples were removed from the temperature-regulating module, gently inverted for 10 s, and analyzed. Repeated-measures models were built to evaluate the effect of time on the stability of ionized Ca (iCa), ionized Mg (iMg), Na, K, Cl, glucose, and lactate. Most of the analytes investigated remained stable up to 8 h under ice water storage conditions before analysis, including iCa, iMg, Cl, glucose, and lactate. However, Na and K were significantly affected by delayed analysis: Na remained stable up to 4 h post-collection, but K was not stable starting at 2 h post-collection. The results of this study are useful in helping future researchers and consultants to recognize acceptable time delays between whole blood collection and processing or analysis for electrolytes, glucose, and lactate.

Liquid storage of dromedary camel semen in different extenders.

This study was conducted to assess the effects of commercial extenders and storage temperature on dromedary camel sperm quality during liquid preservation. In Experiment 1, ejaculates (n = five males; replicated seven times) were split and diluted with synthetic (OPTIXcell, EquiPlus, INRA96, Bioxcell or AndroMed; Experiment 1a) or egg-yolk based (Biladyl, Green buffer or Triladyl; Experiment 1b) extenders and stored for 48 h at 4 °C. In Experiment 2, split ejaculates (n = five males; replicated six times) were used to directly compare Green buffer, OPTIXcell and Triladyl extenders over 48 h of storage at 4 °C. Ejaculates collected in Experiment 3 (n = five males; replicated five times) were diluted with Green buffer or Triladyl before chilled storage for 48 h at 4 or 15 °C. Sperm kinematics, viability and acrosome integrity were assessed during liquid storage. In Experiment 1a, there was the greatest total sperm motility (TM) in the OPTIXcell group following 24 and 48 h of storage, while in Experiment 1b, there was the greatest TM after 48 h of storage with Triladyl and Green buffer. In Experiment 2, there were greater TM and viable acrosome intact spermatozoa in the Triladyl and Green buffer than with OPTIXcell group. In Experiment 3, there was a greater TM in the Triladyl than Green buffer group at 24 and 48 h of storage regardless of storage temperature (which had no effect on sperm quality). In conclusion, camel sperm have greater viability when preserved in liquid form for 48 h following dilution with Triladyl and storage at either 4 or 15 °C.

Causas e prejuízos econômicos por condenação visceral de bovinos em um matadouro frigorífico sob inspeção oficial no município de Corumbá, MS, Brasil / Origin and economic losses decured of bovine visceral condemnation in a slaughterhouse under official inspection in Corumbá, MS, Brazil

O objetivo do estudo foi mensurar as principais causas de condenações viscerais e seus prejuízos econômicos em um matadouro frigorífico localizado na cidade de Corumbá - MS. Foi realizada a quantificação do número de cabeças e órgãos condenados, identificação das principais lesões e estimativa dos prejuízos econômicos causados pelas mesmas. Foram abatidos 21.097 animais, dos quais 13.215 vísceras foram condenadas. Os órgãos com maiores índices de condenações foram os pulmões (47,73%), seguido dos rins (30,2%) e fígados (12,24%). As lesões de maior incidência de condenações foram enfisema pulmonar (27,63%), nefrite (17,43) e aspiração ruminal (13,43%). As condenações levaram a um prejuízo de R$ 73.484,19 em um ano. O fígado foi o órgão responsável por mais da metade das perdas econômicas (56,59%).

Effect of reduced glutathione (GSH) supplementation to Tris-egg yolk extender on chilled semen variables of dogs.

One reason for reduced longevity of chilled dog semen is oxidative stress. The antioxidant glutathione (GSH) improves viability of frozen-thawed dog sperm, but its effect on chilled dog semen has not been investigated. An experiment consisting of two parts was performed: Sperm rich fractions, SRF, were split, diluted with a Tris-egg yolk (TEY) extender containing 0, 5 or 10 mM GSH and stored at 4 °C for 10 days (Part 1; n = 19) or 4 days (Part 2; n = 11), respectively. For Part 1 of the study, percentage (%) of motile, viable, morphologically abnormal spermatozoa and % acrosomal deviations were assessed on days 0, 1, 2, 4 and 10 after dilution. For % sperm motility, samples from all three aliquots of each SRF (0/5/10 mM GSH) were pipetted simultaneously and analysed in a randomised order (time point of analysis, TPA). In Part 2 of the study, motility analysis was performed during 4 days storage and samples were analysed immediately after pipetting (part 2). Most investigated parameters were affected by storage time. For motility variables, there was an effect of GSH identified for circular, CM (ANOVA, Part 1: P = 0.05, Part 2: P < 0.0001) and local motility, LM (ANOVA, Part 2: P = 0.004). Furthermore, there was a trend for an interaction between time and sperm treatment for CM (Part 2: P = 0.077). In conclusion, in the present study there was not an overall positive effect of GSH addition (5/10 mM) on sperm motility in chilled dog semen samples that were characterised to be of good quality during 4- and 10-days of storage.

Effects of oligomeric proanthocyanidins on quality of boar semen during liquid preservation at 17 °C.

The use of antioxidants is an important aspect in the preservation of boar semen. Oligomeric proanthocyanidins (OPC) are an effective natural antioxidants that scavenges free radicals and reactive oxygen species (ROS). This study was designed to investigate the antioxidative effect of OPC on boar semen quality during liquid preservation at 17 °C. The effects of different concentrations of OPC on sperm quality variables, antioxidant effects and fertility were analyzed in this experiment. Semen collected from six Guanzhong-Black boars and was diluted with Beltsville thawing solution (BTS). During the process of liquid preservation at 17 °C, the variables assessed were measured and analyzed every 24 h. The addition of OPC improved boar sperm motility, acrosome integrity, membrane integrity, and mitochondrial membrane potential as compared with that of the control group (P < 0.05). Meanwhile, malondialdehyde content (MDA) and ROS content was less after adding OPC, thereby improving the total antioxidant capacity (T-AOC) (P < 0.05). Different concentrations of OPC have different degrees of protective effects on boar semen quality. The results indicate that 50 µg/mL of OPC was the optimum concentration, and that the conception rate, litter size, and survival rate increased at this concentration as compared with that of the control group (P < 0.05). In summary, the addition of OPC to BTS diluents can improve the quality of boar semen at 17 °C during liquid preservation. Further research is needed to understand the mechanism by which OPC provides protection to boar semen during preservation in vitro.

Effect of refrigeration of clinical canine urine samples on quantitative bacterial culture.

OBJECTIVE To examine the effect of 24 hours of refrigeration on urine samples collected from dogs with signs of urinary tract infection (UTI). DESIGN Prospective cross-sectional study. ANIMALS 104 dogs with signs consistent with UTI that had a urine sample collected via cystocentesis as part of their diagnostic workup. PROCEDURES A 1-mL aliquot of each urine sample was refrigerated at 5°C for 24 hours in a plain glass tube, then processed for quantitative bacterial culture (QBC). A 0.5-mL aliquot was added to 3 mL of tryptic soy broth (TSB) and refrigerated at 5°C for 24 hours, then processed for QBC. The remaining portion was immediately processed for QBC, with results reported as numbers of bacterial colony-forming units (CFUs). Sensitivity of the QBC for detection of bacteria (and therefore UTI) was determined for sample refrigeration in the 2 conditions, compared with immediate processing (reference standard). RESULTS Bacterial growth was identified in 35.6% (n = 37), 33.7% (35), and 31.7% (33) of the immediately processed, refrigerated, and refrigerated-in-TSB urine samples, respectively. Sample refrigeration without TSB resulted in no significant difference in CFU counts relative to immediate processing; however, the sensitivity of this method was 95% (35/37). Sample refrigeration with TSB resulted in significantly lower CFU counts, and sensitivity was only 89% (33/37). CONCLUSIONS AND CLINICAL RELEVANCE Canine urine samples collected for bacterial culture should be immediately submitted for testing. Although CFU counts for refrigerated and immediately processed samples were statistically similar in this study, sample refrigeration in enrichment broth resulted in imperfect sensitivity for UTI detection and is not recommended.

On-farm storage of livestock vaccines may be a risk to vaccine efficacy: a study of the performance of on-farm refrigerators to maintain the correct storage temperature.

BACKGROUND: Livestock vaccines (LV) are often stored on-farm, in a refrigerator (fridge), prior to use and little is documented about the storage conditions during this period. As the quality of a vaccine can be impaired by storage at an incorrect temperature, the present study aimed to evaluate the on-farm performance of farm fridges to maintain the correct storage temperature. From January to August 2014, temperature data loggers were placed on selected farms fridges used to store LV (n = 20) in South-West England. RESULTS: Temperature recording data was available from 17 of the 20 farms. Fifty-nine percent of farm fridges had at least one temperature recording above 8 °C, 53% had at least one recording below 2 °C and 41% at or below 0 °C. Internal fridge temperatures attained 24 °C and dropped to - 12 °C as an absolute maximum and minimum respectively. Fridges tested spent an average of 16% of the total time recorded above 8 °C. Time of the year significantly influenced the percentage of time above 8 °C. External and internal temperatures were found to be positively correlated (p < 0.001). Statistical significant differences in internal and external temperatures were found between March and August. CONCLUSIONS: The majority of fridges in this study would have failed to keep any stored LV within the recommended storage temperature range. If LV are going to be stored on-farm prior to use, then urgent improvements in this part of the cold-chain are required in order to insure vaccine efficacy is not compromised.