Studies on Aspergillus Fumigatus; Toxin Production Ry Different Strains and Serological Comparison of the Strains

Saline extracts of mycelial material from 14 different strains of Aspergillus fumigatus showed haemolytic and toxic activities. These activities were present in filtrates obtained from mycelial material incubated at 20°C for 6, 15 and 30 days and at 37°C for 6 days. The haemolysin and toxin titers decreased with an increase in incubation time and at higher incubation temperatures.The haemolytic and toxic activities could be inactivated by heating the filtrates at 70°C for 10 min.The results of serological tests demonstrated immunological uniformity for the haemolytic and toxic factors.     

The Effects of Cesarean Section Anesthesia on Heat Loss and Heat Production in the Newborn Rabbit

The newborn of some smaller animals rely upon heat produced by nonshivering thermogenesis in the brown fat to prevent a fall in body temperature after birth. Because of their pharmacological properties, some drugs may affect nonshivering thermogenesis. Therefore, in this study, the ability of newborn rabbits delivered under Innovar-Vet, ketamine hydrochloride, methoxyflurane and epidural anesthesia to maintain the rectal, subcutaneous interscapular and lumbar temperature was investigated at an ambient temperature of 35°C or 22°C and the results compared with control newborns delivered without anesthesia. When the newborns were exposed to 35°C, the anesthetics studied had no effect on the ability of the newborn to maintain the rectal temperature and the subcutaneous temperature over the interscapular fat pad was similar to the lumbar subcutaneous temperature thereby indicating that nonshivering thermogenesis was not activated. However, at an ambient temperature of 22°C Innovar-Vet or methoxyflurane reduced the temperature difference between interscapular and lumbar temperatures to 1.6°C compared to 2.5°C in controls and the difference between core temperature and ambient temperature to 3.5°C greater compared to 7.5°C in controls. Ketamine hydrochloride or lidocaine hydrochloride plus meperidine has less effect because these compounds lack adrenergic blocking properties. These data suggest that newborns delivered under anesthetics or tranquillizers that have adrenergic blocking properties require a warm (35°C) environment to prevent a fall in core temperature.     

Distribution of Acid Phosphatase During Autolysis in Bovine Liver

The percentage distribution of acid phosphatase between lysosomes and cytoplasm in bovine liver during autolysis at 37°C was investigated. The share of the cytoplasmic acid phosphatase activity of the total acid phosphatase activity in liver tissue increased during autolysis being before incubation 28–42 % and after 24 hrs.'' incubation at 37°C 63–94 %. Microbiological contamination increased the proportion of cytoplasmic acid phosphatase.When bovine liver was incubated at 37°C for 24 hrs., the activity of the total acid phosphatase decreased to about 50 % of the initial activity. During a 16 days'' incubation at 4°C the total acid phosphatase activity of bovine liver, however, remained unchanged.     

Temperature Related Absorption and Excretion of Sulphadimidine in Rainbow Trout,Salmo Gairdneri

The main purpose of the present study was to see if a kinetic model and a computer program for characterizing the rate of uptake and clearance of chemicals in aquatic organisms (Biofac, OECD’s Chemicals Testing Programme) was applicable to pharmacokinetic studies in fish. The program seems suitable for such studies.The effect of temperature upon rate of absorption of sulphadimidine from medicated water and elimination of the same drug was investigated in rainbow trout. A rise in the temperature from 7° C to 14° C more than doubled the calculated rate constants of absorption and elimination. The biological half life (t_½ β) and the time to reach 90 % of steady state (t_{90% ss}) at 14° C were approximately half the values at 7° C.     

Comparison of Four Enrichment Media in the Recovery of Campyloracter Jejuni

The effectiveness of 4 enrichment media for the recovery of low levels of inoculated cells of Campylobacter jejuni was evaluated. The media contained antibiotics or antibiotics and bile acids as selective compounds. Three of the media recovered most of the inoculated low numbers of 6 C. jejuni strains. In the 3 media the growth rate of 3 strains, indicated by the increase in the log number of cells during 24 h or 48 h incubation at 42 ° C, was about the same as in the control medium without selective compounds. The same 3 media also recovered a low number of Campylobacter cells from artificially contaminated raw milk or ground meat samples. The enrichment medium B containing 40 I.U. Colistin, 5 μg novobiocin, 2 mg Na-cholic acid and 50 mg cycloheximide per ml was inhibitory for most Campylobacter strains studied.     

Effect of storage time and temperature on the total protein concentration and electrophoretic fractions in equine serum

Serum protein electrophoresis (SPE) is a technique that could be considered one of the most useful diagnostic aids available to the clinician. The effect of storage time and temperature on the total proteins and electrophoretic fractions (albumin, α₁-, α₂-, β₁-, β₂-, and γ-globulins) was assessed in 24 healthy horses. All samples, collected by jugular vein puncture, were centrifuged and divided into 4 aliquots. The 1st aliquot was analyzed within 3 h from collection (time 0), the 2nd was refrigerated at +4°C for 24 h, the 3rd was refrigerated at +4°C for 48 h, and the last was frozen at −20°C for 48 h. One-way repeated-measures analysis of variance (ANOVA) showed a significant effect (P < 0.05) of the different storage conditions on the concentrations of all the parameters studied and significant variations in the percentages of albumin, α₁-globulins, α₂-globulins, and γ-globulins. Compared with time 0 the total protein concentration increased significantly after 48 h at −20°C, the albumin percentage decreased after 48 h at −20°C, the α₁-globulin percentage increased after 24 h at +4°C, the α₂-globulin percentage increased after 48 h at +4°C and at −20°C, and the γ-globulin percentage increased after 48 h at −20°C. The results should help veterinary practitioners handle and store equine serum samples appropriately. Further investigations at different storage times and temperatures could be useful.     

Effect of Recovery Medium on the Isolation of Campylobacter Jejuni Before and After Heat Treatment

The effect of various concentrations of polymyxin B and Colistin in Skirrow’s or Butzler’s type media, respectively, on the recovery rate of 13 strains of G. jejuni before and after heat treatment was studied. Prior to heating, a Butz-ler’s type medium containing 40 I.U. per ml Colistin was inhibitory for certain strains of C. jejuni. After heating at 48° G for 30 min, media containing 2.5 I.U. per ml polymyxin B or 10 I.U. per ml Colistin were not inhibitory for heat-injured cells. When the concentrations of polymyxin B were increased to 5.0 I.U. per ml or those of Colistin to 20 or 40 I.U. per ml in the selective media, the means of the differences of log cell counts between non-selective Brucella blood agar and the selective media was statistically significant (P < 0.05). The mean D-value of G. jejuni strains in Brucella broth at 48° G was 18.4±5.4 min.     

Establishing conditions for the storage and elution of rabies virus RNA using FTA? cards

The Flinders Technology Associates filter paper cards (FTA^® cards) can be used to store nucleic acid from various samples and are easily portable. However, RNA is physicochemically unstable compared with DNA, and appropriate methods have not been established for storage and extraction of RNA from FTA^® cards. The present study investigated the optimum conditions for storage and elution of viral RNA (vRNA) using rabies virus (RABV) applied to FTA^® cards. When TE buffer was used, the elution rates of vRNA increased with the length of the elution time. When the cards were stored at −80°C or −20°C, vRNA was stable over 3 months. Degradation of vRNAs occurred following storage at 4°C and room temperature, suggesting that RNA should be extracted from cards as soon as possible if no freezer is available. When we tried to amplify vRNA from RABV-infected animal brains applied to FTA^® cards and stored at −80°C for 6 months, we did not detect any amplified products with the primer set for 964 bp of RABV N gene. However, we were able to detect amplified products by increasing the elution time of vRNA from FTA^® cards from 30 min to 24 hr or by changing the primer sets to amplify 290 bp of N gene. Thus, we recommend extending the elution time for damaged or low concentration samples in FTA^® cards.     

Procedure for Blood Glutathione Peroxidase Determination in Cattle and Swine

An improved testing system has been developed for direct measurement of glutathione peroxidase activity in heparinized whole blood at 37°C. Without loss in net yield the consumption of reagents has been found to be considerably lower with the new technique than with previously described techniques. Within the range 0–700 mKat/1 the GSH-Px activity in red cells may be measured with a high degree of accuracy and reproducibility without preceding separation and washing. The stability of the enzyme in bovine and porcine whole blood at 22°C, 4°C, and —20°C was determined.     

Comparison between core temperatures measured telemetrically using the CorTemp? ingestible temperature sensor and rectal temperature in healthy Labrador retrievers

This study evaluated the CorTemp^® ingestible telemetric core body temperature sensor in dogs, to establish the relationship between rectal temperature and telemetrically measured core body temperature at rest and during exercise, and to examine the effect of sensor location in the gastrointestinal (GI) tract on measured core temperature. CorTemp^® sensors were administered orally to fasted Labrador retriever dogs and radiographs were taken to document sensor location. Core and rectal temperatures were monitored throughout the day in 6 resting dogs and during a 10-minute strenuous retrieving exercise in 6 dogs. Time required for the sensor to leave the stomach (120 to 610 min) was variable. Measured core temperature was consistently higher than rectal temperature across all GI locations but temperature differences based on GI location were not significant (P = 0.5218). Resting dogs had a core temperature that was on average 0.4°C above their rectal temperature with 95% limits of agreement (LoA) between 1.2°C and −0.5°C. Core temperature in exercising dogs was on average 0.3°C higher than their concurrent rectal temperature, with LoA of +1.6°C and −1.1°C.     

The effect of breed,sex, and oral meloxicam administration on pain biomarkers following hot-iron branding in Hereford and Angus calves

Hot-iron branding uses thermal injury to permanently identify cattle causing painful tissue damage. The primary objective was to examine the physiological and behavioral effects of oral meloxicam (MEL), compared to a control, administered at the time of hot-iron branding in Angus and Hereford steers and heifers. The secondary objectives were to investigate breed and sex effects on pain biomarkers. A total of 70 yearlings, consisting of 35 heifers and 35 steers (Angus, Hereford, or Angus × Hereford), were enrolled in the study. Animals were blocked by sex, randomized across weight, and assigned to receive MEL (1 mg/kg) or a placebo (CON). Biomarkers were assessed for 48 h after branding and included infrared thermography (IRT), mechanical nociceptive threshold (MNT), accelerometry and a visual analog scale (VAS), and serum cortisol and prostaglandin E₂ metabolites (PGEM). Wound healing was assessed for 12 wk. Hair samples to quantify cortisol levels were taken prior to and 30 d post-branding. Responses were analyzed using repeated measures with calf nested in treatment as a random effect, and treatment, time, treatment by time interaction, breed, and sex as fixed effects. There was a treatment by time interaction for PGEM (P < 0.01) with MEL having lower values than CON at 6, 24, and 48 h (MEL: 18.34 ± 3.52, 19.61 ± 3.48, and 22.24 ± 3.48 pg/mL, respectively; CON: 32.57 ± 3.58, 37.00 ± 3.52, and 33.07 ± 3.48 pg/mL; P < 0.01). MEL showed less of a difference in maximum IRT values between the branded (2.27 ± 0.29 °C) and control site (3.15 ± 0.29 °C; P < 0.01). MEL took fewer lying bouts at 0–12 h (4.91 bouts ± 0.56) compared with CON (6.87 bouts ± 0.55; P < 0.01). Compared with Hereford calves, Angus calves exhibited greater serum but lower hair cortisol, greater PGEM, more lying bouts, and less healed wound scores at 3, 4, and 5 wk. Compared with heifers, steers exhibited lower PGEM, lower branding site and ocular IRT, higher MNT, and lower plasma meloxicam levels. Steers spent more time lying, took more lying bouts and had greater VAS pain, and more healed wound scores at 5 wk than heifers. Meloxicam administration at branding reduced branding and control site temperature differences and reduced lying bouts for the first 12 h. Breed and sex effects were observed across many biomarkers. Changes from baseline values for IRT, MNT, lying time, step count, VAS pain, and wound scoring all support that branding cattle is painful.     

Determination of the best practical method of thawing bovine semen

An experiment was designed to determine the best practical method of thawing which could be applied to all semen processed by Canadian A.I. centers. Semen in Tris, Triladyl, whole milk, or Fresh Plus Extender packaged in 1 mL ampules or 0.5 mL French straws was used. Semen was thawed in water at 35°C, 20°C, or 5°C, or in a shirt pocket for 5, 5, 10, or 10 minutes, respectively. Post-thaw viability of sperm cells was assessed by determining the percentage of progressively motile cells, rate of progression, and percentage of intact acrosomes, at 0 hours and after 2 hours of incubation at 37°C. Post-thaw viability was significantly higher (P < 0.05) for the 35°C thaw than for any of the other thaw methods regardless of extender or packaging. Our data indicate that when AI center recommendations for thawing semen are unknown, semen in ampules and straws should be thawed in a 35°C water bath and maintained at that temperature until it can be inseminated.     

The Effect of Customary Finnish Heat Preparation Methods on the Infestiveness of Diphyllobothrium Latum From Fish to Man an Experimental Treatise on the Heat Resistance of Diphyllobothrium Latum Plerocercoids,and on the Temperature Variations in Various Parts of the Pike,Perch and Burbot During Frying in Pan,Boiling, and Baking in Oven

In part A of the investigation the heat resistance of isolated Diphyllobothrium plerocercoids was determined. Treatments for 5 min/at 56°C, 10 min. at 50°C, 20 min. at 48°C, and 30 min. at 47°C were found to be lethal as indicated by cessation of spontaneous movements. The number of plerocercoids in the tests was 640.In part B the consumption of pike, perch and burbot, which are known to be most important carriers of D. latum plerocercoids, and the customary methods of preparation of these fish species in Finland are first reviewed. Then the results of 85 frying and 7 boiling experiments are described, in which the temperature change at the sites of the least temperature rise was followed. During frying in the pan the inner temperature of the fish rose to the critical 56° C limit during 10 to 20 min., depending on the size of the fish. Normal boiling and frying in an oven were found to be rather safe in this respect.In all tests fish heated to the critical temperature of the plerocercoids were organoleptically found to be ready for the table.     

Appraisal of Collection Techniques and Storage Temperatures on Turkey Plasma Cholinesterase Levels and Blood Glutathione Concentrations

The effects of different blood collection procedures, various storage temperatures and durations of storage on the levels of plasma cholinesterase and whole blood glutathione in turkeys were investigated.

Collection of blood through vacutainers yielded satisfactory results. Whereas the plasma cholinesterase activity remained unchanged even after three weeks of storage at -17.8°C., blood glutathione concentration was unaffected only when the samples were stored at -28.9°C for the three weeks. The range of mean activity was from 4.64 to 4.71 ΔpH/hour x 10 for cholinesterase and from 44.85 to 47.91 mgm/100 ml for glutathione.

     

Spermine protects intestinal barrier integrity through ras-related C3 botulinum toxin substrate 1/phospholipase C-γ1 signaling pathway in piglets

Weaning stress can cause tight junctions damage and intestinal permeability enhancement, which leads to intestinal imbalance and growth retardation, thereby causing damage to piglet growth and development. Spermine can reduce stress. However, the mechanism of spermine modulating the intestinal integrity in pigs remains largely unknown. This study aims to examine whether spermine protects the intestinal barrier integrity of piglets through ras-related C3 botulinum toxin substrate 1 (Rac1)/phospholipase C-γ1 (PLC-γ1) signaling pathway. In vivo, 80 piglets were categorised into 4 control groups and 4 spermine groups (10 piglets per group). The piglets were fed with normal saline or spermine at 0.4 mmol/kg BW for 7 h and 3, 6 and 9 d. In vitro, we investigated whether spermine protects the intestinal barrier after a tumor necrosis factor α (TNF-α) challenge through Rac1/PLC-γ1 signaling pathway. The in vivo study found that spermine supplementation increased tight junction protein mRNA levels and Rac1/PLC-γ1 signaling pathway gene expression in the jejunum of piglets. The serum D-lactate content was significantly decreased after spermine supplementation (P < 0.05). The in vitro study found that 0.1 μmol/L spermine increased the levels of tight junction protein expression, Rac1/PLC-γ1 signaling pathway and transepithelial electrical resistance, and decreased paracellular permeability (P < 0.05). Further experiments demonstrated that spermine supplementation enhanced the levels of tight junction protein expression, Rac1/PLC-γ1 signaling pathway and transepithelial electrical resistance, and decreased paracellular permeability compared with the NSC-23766 and U73122 treatment with spermine after TNF-α challenge (P < 0.05). Collectively, spermine protects intestinal barrier integrity through Rac1/PLC-γ1 signaling pathway in piglets.     

In utero heat stress alters the postnatal innate immune response of pigs

The effects of in utero heat stress (IUHS) range from decreased growth performance to altered behavior, but the long-term impact of IUHS on postnatal innate immune function in pigs is unknown. Therefore, the study objective was to determine the effects of early gestation IUHS on the immune, metabolic, and stress response of pigs subjected to an 8 hr lipopolysaccharide (LPS) challenge during postnatal life. Twenty-four pregnant gilts were exposed to thermoneutral (TN; n = 12; 17.5 ± 2.1 °C) or heat stress (HS; n = 12; cyclic 26 to 36 °C) conditions from days 6 to 59 of gestation, and then TN conditions (20.9 ± 2.3 °C) from day 60 of gestation to farrowing. At 12 wk of age, 16 IUHS and 16 in utero thermoneutral (IUTN) pigs were selected, balanced by sex and given an intravenous injection of LPS (2 µg/kg BW mixed with sterile saline [SAL] and injected at 2 µL/kg BW) or SAL (2 µL/kg BW). Body temperature was monitored every 30 min, and blood was obtained at 0, 1, 2, 3, 4, 6, and 8 hr following the LPS challenge. Blood samples were analyzed for glucose, insulin, non-esterified fatty acids (NEFA), cortisol, and cytokine concentrations. In addition, white blood cell counts were determined at 0 and 4 hr. Hour 0 data were used as covariates. Body temperature was increased (P < 0.01) in LPS (40.88 ± 0.08 °C) vs. SAL (39.83 ± 0.08 °C) pigs. Eosinophils tended to be decreased overall (P = 0.09; 43.9%) in IUHS vs. IUTN pigs. Glucose concentrations were reduced overall (P = 0.05; 5.9%) in IUHS vs. IUTN pigs. The NEFA concentrations tended to be greater (P = 0.07; 143.4%) in IUHS-LPS pigs compared with all other treatments, and IUTN-LPS pigs tended to have greater (127.4%) circulating NEFA concentrations compared with IUTN-SAL and IUHS-SAL pigs. Cortisol was increased (P = 0.04) in IUHS-LPS compared with IUTN-LPS pigs at 3 hr (21.5%) and 4 hr (64.3%). At 1 hr, tumor necrosis factor α was increased (P = 0.01; 115.1%) in IUHS-LPS compared with IUTN-LPS pigs. Overall, interleukin-1β (IL-1β) and interleukin-6 (IL-6) were greater (P < 0.04; 281.3% and 297.8%, respectively) in IUHS-LPS pigs compared with all other treatments, and IUTN-LPS pigs had increased IL-1β and IL-6 concentrations compared with IUTN-SAL and IUHS-SAL pigs. In summary, IUHS altered the postnatal cytokine, metabolic, and physiological stress response of pigs during postnatal life, which may have negative implications toward the innate immune response of IUHS pigs to pathogens.     

Assessment of stability of ketamine-xylazine preparations with or without acepromazine using high performance liquid chromatography-mass spectrometry

The objective of this study was to evaluate the stability of 3 distinct preparations of ketamine and xylazine, with or without acepromazine, stored at room temperature or at 4°C for 1, 2, and 3 mo. Drug concentrations were compared to fresh solutions, using a high performance liquid chromatography-mass spectrometry/selected-ion monitoring (HPLC-MS/SIM) assay. The concentrations of ketamine and xylazine, diluted in physiological saline, did not change over time at room temperature or at 4°C. However, acepromazine concentrations decreased over time when stored at room temperature. In contrast, undiluted ketamine-xylazine preparations gradually decreased in concentration when stored at room temperature. All of the drug concentrations remained above 90% of their original concentration when stored at 4°C. In conclusion, when diluted in physiological saline, ketamine-xylazine cocktails can be stored for 3 mo, whereas undiluted cocktails can lose efficacy over 3 mo at room temperature. Storage at 4°C could preserve drug stability.     

Association between in-transit loss, internal trailer temperature, and distance traveled by Ontario market hogs

An observational study was conducted from July to October 2004 to determine the association between in-transit losses of swine and internal trailer temperature after controlling for loading density, trip distance, herd size, and random trip effect. A convenience sample of 3 trucking companies was used to collect temperature, relative humidity, and global positioning data for 104 trips that delivered 21 834 pigs from 371 producers to Ontario abattoirs. The association between in-transit loss and trailer temperature was determined using the 90th percentiles of internal temperature for each trip. Average loading density was 0.36 m²/100 kg pig (range 0.28 to 0.50 m²/100 kg pig). Average in-transit loss was 0.12%; however, 94% of producers experienced no losses. As the 90th percentile of internal trailer temperature increased from a range of 8.6°C to 23.3°C to a range of 23.4°C to 26.1°C, average in-transit loss ratio increased approximately 3-fold, with an additional 2-fold increase as the range increased from 26.2°C to 28.9°C to 29.0°C to 30.5°C. As the 90th percentile of temperature increased by 1°C over the full range of temperatures in this study, in-transit loss was expected to increase 1.26 times. The in-transit loss was expected to decrease 0.81 times for each 50-km increase in distance traveled between the farm and the abattoir.     

Survival of thermotolerant campylobacters in water

Three bacterial strains, classified as Campylobacter jejuni biotype 1, Campylobacter coli, and NARTG (nalidixic-acid-resistant thermophilic Campylobacters), were tested for survival in water specimens kept at 4, 12, and 20°C. Five different water milieus were compared: sterile physiological saline, chlorinated tap water, dechlorinated tap water, polluted river water, and sterile filtered river water. With few exceptions, all organisms survived better at 4°C than at 12 or 20°C, regardless of the water milieu. Briefest survival was detected at 20°C; no viable Campylobacters could be demonstrated after more than 2 days at this temperature. Of the 5 waiter milieus tested, the highest mean survival time for all strains was obtained with dechlorinated tap water. In this medium, all 3 strains remained viable for 15 days at 4°C, 10 days at 12°C, and 2 days ait 20°C. Briefest survival was obtained in chlorinated tap water. Even residual amounts of Cl₂ drastically reduced the survival of all strains tested. Only small variations in viability were detected for 2 of the strains tested after sterile filtration of a water source with a dense bacterial population. The results are discussed in relation to waterborne outbreaks of campylobacteriosis.     

A Comparison of Seitz and Millipore Membrane Filters for the Enumeration of Fecal Coliforms

In a comparison of two commonly used membrane filters for enumerating fecal coliform bacteria it was demonstrated that Seitz type M filters recovered statistically more colonies of bacteria than did Millipore HAWG 047S1 filters from pure cultures of Escherichia coli incubated at 44 °C. The membranes were grown on 0.4 % Teepol agar. On incubation at 37°C no significant discrepancy was found. As a reference method was used pour plating in plate count agar (Difco). It was demonstrated that incubation at 44°C did not per se inhibit propagation of fecal coliforms. Both types of filters examined were sterilized by the manufacturers with ethylene oxide. The discrepancy found can therefore not be due to sterilization procedures.