Stimulation of the titre of a sheep serum factor that is related to the sheep complement system

Serum from most sheep subjected to a single jugular bleeding, repeated bleeding or an intraperitoneal injection of yeast and repeated bleeding, showed an increase in the titre of a serum component called serum factor. Serum factor reached a peak titre 2 to 5 days after treatment started. For some sheep, the titre was elevated for a 9- to 12-day period whereas for others the titre dropped markedly on day 7 followed by a rise on day 9. Serum factor reacts with sheep erythrocytes sensitised with rabbit antibody (sheep E-rabbit A). Serum factor can be detected on sheep E-rabbit A using guinea-pig antiserum that reacts with sheep complement (C). Serum factor is inactivated by heating at 56 degrees C for 30 minutes, but is only partially inactivated at 50 degrees C for 30 minutes. The reaction of serum factor with sheep E-rabbit A is inhibited by chelators of Ca++ and/or Mg++. Serum factor appears to be related to the sheep C system. Preliminary results suggest it may he a component of the classical pathway of sheep C, possibly the second component, C2.     

Antibacterial spectrum and some other characteristics of an antimicrobial factor produced by Yersinia ruckeri

Yersinia ruckeri produces an antibacterial factor which inhibits the growth of a wide spectrum of Gram-negative and Gram-positive bacteria, though not other strains of Y. ruckeri. The antibacterial factor was produced at low temperatures (4-20 degrees C), but not at 37 degrees C. The activity was lost after treatment of the supernatant with chloroform, UV-light and after boiling of the supernatant. One did not succeed in obtaining the antibacterial factor in a sterile solution.     

Factors affecting the survival of Streptococcus suis type 2

The survival of Streptococcus suis type 2 was assessed in experimentally inoculated faeces and dust stored at 0, 9 and 22 to 25 degrees C. The organism survived in faeces for 104 days at 0 degrees C, up to 10 days at 9 degrees C and up to eight days at 22 to 25 degrees C. It survived in dust for up to 54 days at 0 degrees C and up to 25 days at 9 degrees C but could not be isolated from dust stored at room temperature for 24 hours. The organism survived at 4 degrees C in nutrient medium for up to nine months but in distilled water for only one to two weeks. At 50 degrees C it survived in water or broth for up to two hours but at 60 degrees C it only survived for 10 minutes. The organism was rapidly inactivated by disinfectants and cleansers, commonly used on farms and in laboratories, at concentrations less than those recommended for use by the manufacturers.     

Alternative pathway fof bovine complement Immunochemical studies on factor B-like serum protein and its conversion product B gamma 2.

Rabbits produced antibodies to a factor B-like serum protein (factor Bbov), its conversion product B gamma 2 and some other bovine serum proteins after repeated immunization with zymosan which previously had been incubated with fresh bovine serum. Such antisera were used to monitor purification of B gamma 2 from fresh bovine sera incubated with zxymosan. Subsequently, antisera specific for factor Bbov and B gamma 2 were produced. Antiserum produced against B gamma 2 cross-reacted with factor Bbov. Functional assays for factor Bbov were carried out in a hemolytic system with guinea pig erythrocytes in EGTA buffer. Heat inactivation (56 degrees C/5 min) of bovine serum destroyed the antigenicity of factor Bbov but not that of B gamma 2. Factor Bbov had an apparent molecular weight of 95,000 and B gamma 2 a molecular weight of 40,000 daltons. Conversion of factor Bbov to B gamma 2 was determined qualitatively by immunoelectrophoresis and quantitatively by radial immunodiffusion. Conversion of factor Bbov to B gamma 2 in bovine serum, in the presence of zymosan or cobra venom factor (CoVF) required Mg++ but not Ca++, did not occur in heat inactivated (56 degrees C/5 min) serum and was maximal, but not complete, when fresh bovine serum was incubated with zymosan (20 mg/mL) at 37 for two hours.     

Stability of canine factor VIII activity and von Willebrand factor antigen concentration in vitro

The in vitro stability of canine factor VIII activity, von Willebrand factor antigen concentration and the ratio of these two factors was studied. Samples were stored for up to 48 hours, either as plasma or as whole blood, at 4 degrees, 20 degrees and 37 degrees C. Factor VIII activity was generally stable in both plasma and whole blood samples for up to 48 hours at 4 degrees or 20 degrees C. The concentration of von Willebrand factor antigen was more stable in samples stored as plasma than whole blood, and for a shorter time than factor VIII activity. Consequently, the stability of the ratio of these two factors was relatively poor in vitro.     

Studies on Rhinoestrus purpureus (Diptera: Oestridae) larvae infesting donkeys (Equus asinus) in Egypt. III. Pupal duration under controlled conditions.

The pupal duration of Rhinoestrus purpureus was studied under variable degrees of temperature and relative humidity (RH). It was found that pupal duration was affected by temperature but not by RH. An increase in the temperature above 22 degrees C decreased the pupal duration: 26-27 days at 22 degrees C, 16-24 days at 27 degrees C, 13-15 days at 32 degrees C. At 37 degrees C, the pupated larvae failed to pupate and died. The deformity ratio of emerged flies was 25-30% at 22-32 degrees C, but it was directly proportional to RH at a constant temperature of 32 degrees C: 26.1% at 75% RH, 16.7% at 50% RH, 6.7% at 30% RH. It was concluded that the optimum temperature and RH for obtaining normal active flies were 32 degrees C and 30%, respectively.     

Analysis of virulence plasmid gene expression of intra-macrophage and in vitro grown Rhodococcus equi ATCC 33701

Rhodococcus equi is a soil organism that infects macrophages of foals and immunocompromised humans. Virulence in foal isolates is tightly associated with an 80kb plasmid, which includes a pathogenicity island (PI) with a virulence-associated gene family, vap. A DNA microarray containing 66 of 69 putative open reading frames (ORFs) of the virulence plasmid was developed. Virulence plasmid gene expression of R. equi grown in macrophages or under different conditions in vitro was compared against in vitro growth at 30 degrees C, pH=7. When grown in macrophages, all seven vap family genes as well as six ORFs within, but not outside, the PI were induced. Cluster analysis of the gene expression matrix assembled from different growth conditions suggested that those genes that actively responded to environmental changes divided broadly into two groups. One group, orf1, 2, 5, 6-8, 12-15, 19, and 20 (which includes all the vap genes), was induced at 37 degrees C, mostly by low iron, and to a lesser extent by the synergy of low calcium and pH=5. The second group, orf3, 9, and 10, was induced at 37 degrees C by magnesium depletion (produced by EDTA treatment of growth medium). Temperature (37 degrees C) was the most important factor inducing gene expression for the both groups. Iron restriction led to down-regulation of Group II genes and magnesium restriction led to down-regulation of Group I genes. A putative consensus IdeR binding site was identified upstream of vapA, suggesting that vapA is a member of an IdeR regulon in R. equi. Expression of genes inside macrophages was most closely but not completely mimicked by growth of bacteria at 37 degrees C, pH=5, under conditions of restricted iron, calcium and magnesium; that is, similar to environmental factors found inside macrophages.     

Cytotoxin from an ovine strain of Pasteurella haemolytica: characterisation studies and partial purification

A cell-free, water-soluble cytotoxin from an ovine strain of Pasteurella haemolytica biotype A serotype 1 killed sheep bronchoalveolar macrophages at 37 degrees C, but not at 4 degrees C or 22 degrees C. The cytotoxin was stable over the pH range 2-12, resistant to heat at 60 degrees C but inactivated at 100 degrees C or by autoclaving. Trypsin also destroyed the cytotoxin, which is therefore thought to contain a protein component essential for biological activity. A preliminary purification of the crude cytotoxin using gel-filtration column chromatography resulted in the isolation of a biologically active fraction which resolved as a single protein band and one carbohydrate band on non-dissociating polyacrylamide gels. However, this fraction resolved into approximately 16 component bands on a sodium dodecyl sulphate polyacrylamide gel.     

Effects of ambient temperature on heat increment of feeding and energy retention in growing broilers maintained at different food intakes

Zero-activity heat production (HP), body temperature (Tb) and energy retention were measured in growing broilers maintained at 5 ambient temperatures (Ta) (14 degrees , 17 degrees , 22 degrees , 27 degrees and 32 degrees C) and at 5 feeding rates (ad libitum intake and 75%, 50%, 25% and 0% (fasting) of ad libitum). Zero-activity HP increased with decreasing Ta and increasing food intake. However, at 14 degrees C, zero-activity HP in birds fed ad libitum and 75% did not show further increase, but those in birds fed less than 75% of ad libitum increased rapidly. Results of the regression of zero-activity HP on Ta ranging from 32 degrees to 17 degrees C indicated that the slope was affected little by food intake, but the intercept decreased with decreasing food intake. Tb increased significantly with increasing food intake. There was little variation with Ta but, at and above 27 degrees C, a slightly increased Tb was observed only in birds fed ad libitum. Overall effects of Ta and food intake on HIF (% TME intake) were not found, but HIF tended to increase with decreasing food intake at 14 degrees C. Total energy retention and energy retention as fat decreased with decreasing Ta and food intake, although energy retention as protein decreased only with decreasing food intake. Results obtained here suggest that availability of TME is affected little by Ta ranging from 32 degrees to 17 degrees C and that HIF is utilised, in part, to maintain Tb at any Ta.     

Effects of constant environmental temperatures on the performance of laying pullets

1. Two experiments are described in which laying pullets maintained at constant temperatures were fed a range of diets with a view to defining optimum combinations of temperature and nutrient intake. 2. In the first experiment, all combinations of 6 temperatures (15 degrees, 18 degrees, 21 degrees, 24 degrees, 27 degrees and 30 degrees C) 9 diets (three protein concentrations and three energy contents) and two stocks were tested for 34 weeks using 4320 pullets. In experiment 2, all combinations of three rearing temperatures, three laying temperatures (18 degrees, 22.5 degrees and 27 degrees C) three diets (protein concentration) and two stocks were tested for 61 weeks using 2160 pullets. 3. As anticipated, higher dietary protein concentrations were needed to maintain egg output at higher temperatures. If diets suplying adequate amino acid intakes were provided, egg output was unaffected by temperatures in the range 15 degrees to 27 degrees C although, at the highest temperature, egg weight was slightly reduced and rate of lay (particularly in the later part of the laying year) was increased. At 30 degrees C, egg output was depressed whichever diet was fed. 4. Dietary energy content had small but significant effects on egg weight and egg output but did not interact with temperature. It was not possible to maintain egg weight or egg output at 30 degrees C by feeding a high energy, high protein diet. 5. Estimated heat output of the birds increased during the course of the experiment at the lower temperatures but decreased with time at 30 degrees C. Feather loss occurred earlier at the lower temperatures and this is interpreted as an effect of temperature on the timing of the annual moult, which also accounts for the better persistency of lay observed at 27 degrees C.     

Activity of supplemental enzymes and their effect on nutrient utilization and growth performance of growing chickens as affected by pelleting temperature.

Activity of supplemental enzymes in a barley-soybean-maize based diet at 60, 75 and 90 degrees C pelleting temperatures was studied using feed viscosity, in-vitro enzyme activity and broiler performance data. High pelleting temperatures increased feed viscosity but supplemented enzymes reduced the viscosity at all three temperatures levels by 11, 14 and 17%, respectively. Water intake and losses in excreta of birds were found to be affected by feed viscosity. Activity of cellulase enzyme, measured using the radial diffusion method, was unaffected at 60 and 75 degrees C, but reduced by 73% in feed processed at 90 degrees C. Enzymes increased the weight gain of broilers by 11.1% at 90 degrees C, but no effect could be seen at low pelleting temperatures possibly due to high dietary protein and energy contents. Feed intake was unaffected by enzymes. Birds consumed 6% more feed and grew 9% faster when the pelleting temperature was increased from 60 to 75 degrees C. Reduced feed intake and daily weight gain observed at 90 degrees C could be fully compensated by the enzyme supplementation. High pelleting temperature reduced energy metabolizability (3.2%) and nitrogen utilization (4%) but enzyme almost compensated them (by 3.3% and 2.6%, respectively). No interaction could be detected between the pelleting temperatures and enzymes. It is concluded that pelleting temperatures as high as 90 degrees C drastically reduce cellulase activity, energy and nitrogen utilization thus lowering broiler performance. Either the remaining activity of cellulase or other thermostable enzymes can prevent the losses.     

Factors contributing to the resistance of chickens to infection with Japanese Fasciola sp.

Attempts were made to clarify the factors contributing to the resistance of chickens to infection with Japanese Fasciola sp. Infection was not successfully established in chickens by oral inoculation of metacercariae, nor by inoculation of excysted juvenile flukes into the body cavity or to the liver surface. Many metacercarial cysts were detected within two days in the feces of orally inoculated chickens. In the in vitro excystation test with chicken bile at 42 degrees C, metacercariae emerged successfully. These results indicate that the major resistant factors may not act during the migration from the mouth to the liver. Histopathological examination of the liver of experimental chickens could not prove the effect of a resistant factor. Excysted flukes were cultivated at 37-42 degrees C in RPMI1640 supplemented with calf serum, with the result that the survival rate of flukes fell with higher temperatures. When chicken serum was used instead of calf serum, flukes survived for a long period of time at 37 degrees C, while all died within four days at 42 degrees C. The higher body temperature of chickens than that of other mammalian hosts is considered to be the major factor contributing to the resistance of chickens to infection with Fasciola sp.     

Effect of a change in environmental temperature on heat tolerance in laying fowl

Laying hens when transferred from accommodation at an ambient temperature (Ta) of 30 degrees C to one of 20 degrees C failed to acclimatise to intermittent heat stress (Ta 38 degrees C) commencing one day after the transfer. After 21 d of intermittent exposure to 38 degrees C these hens showed little or no increase in heat tolerance, whereas hens living constantly at either Ta 20 degrees C or 30 degrees C acclimatised normally. The failure to acclimatise was also observed when hens were transferred from Ta 30 degrees C to 5 degrees C but not when transferred from Ta 5 degrees C to 30 degrees C. The failure to acclimatise following transfer from a warm to a cool environment was accompanied by an increase in food intake; if food intake was not allowed to increase the hens acclimatised normally to heat stress.     

Effect of infection by Babesia spp. on the development and survival of free-living stages of Boophilus annulatus

Oviposition, egg hatching and survival of newly-hatched larvae of Boophilus annulatus were studied in relation to infection by Babesia species and different temperature regimens. Infection of female ticks by Babesia bigemina or B. bovis had no effect on the time elapsed between engorgement and oviposition. The duration of oviposition was shorter in infected females incubated at 25 degrees C or 35 degrees C and infected females laid fewer eggs than the controls. No larvae hatched at 16 degrees C. B. bigemina-infected eggs hatched more quickly than uninfected eggs at 35 degrees C. The hatching percentage of B. bigemina-infected eggs was reduced by 50% at an incubation temperature of 25 degrees C and by 75% at 35 degrees C. At 16 degrees C there was no difference in the duration of survival of infected and non-infected larvae but at 25 degrees C and 35 degrees C the mean survival period of infected larvae was significantly lower than those of controls.     

Some effects of temperature on the adults, eggs and pupae of Stomoxys calcitrans Linnaeus (Diptera: Muscidae)

Adults could only live and reproduce to their full capacity at temperatures between 20 degrees C and 30 degrees C. At 15 degrees C the females laid no eggs, the adult life span was relatively short and the reproductive capacity of females kept at 35 degrees C was low. The thermal histories of the flies had no apparent effect on their later reactions to temperature in any of the parameters tested. The viability rates of S. calcitrans eggs exposed to temperatures between 10 degrees C and 40 degrees C exceeded 84%, but 45 degrees C was lethal. The optimum temperatures for incubation of the eggs was 30 degrees C. Pupae of S. calcitrans seemed to tolerate temperatures between 20 degrees C and 30 degrees C, but their mortalities increased markedly outside this temperature range. Tests showed that pupal mortalities increased linearly with increasing periods of exposure to a temperature of 15 degrees C.     

Molecular pathogenesis of equine coital exanthema: temperature-sensitive function(s) in cells infected with equine herpesviruses

Preliminary experiments have revealed that several laboratory and wild-type strains of the equine herpesvirus (EHV) triad were temperature-sensitive for growth when assayed at 39 degrees C. The efficiencies of plating (EOP) observed were 10(-2) for both EHV 1 and 2, and 1 X 10(-6) for EHV 3. The EOPs were determined by plaque assays which compared titrations at 34 degrees C and 39 degrees C on equine fetal dermal fibroblast cells. Growth yield experiments, assayed at 34 degrees C, reflected those EOP's, but did not indicate any difference in yields when infected cultures were incubated at 34 degrees C and 37 degrees C. Temperature shift experiments with EHV 3-infected cultures revealed that a temperature-sensitive function(s) responsible for the reduction in titer appeared to be a late function(s). All strains examined appeared to incorporate H3-thymidine into viral-density DNA at the non-permissive temperature of 39 degrees C. Electron microscopy of EHV 3-infected cell cultures, incubated continuously at the non-permissive temperature and examined at 18 h after infection, revealed structures consistent with the accumulation of nucleocapsids within the nucleus. The evidence presented is consistent with the hypothesis that in equine dermal cells infected with a plaque-purified wild-type strain of EHV 3 (1118LP), a function needed for the egress of nucleocapsids from the nucleus is absent at 39 degrees C. The significance of these findings relative to the pathogenicity of the disease (equine coital exanthema) caused by this virus is discussed.     

Effects of heating and freezing on the viability of sarcocysts of Sarcocystis levinei from cardiac tissues of buffaloes

Dogs fed buffalo heart muscle containing sarcocysts of Sarcosystis levinei and heated at 65-75 degrees C did not shed sporocysts, whereas other dogs fed infected heart muscle heated between 40 and 60 degrees C shed sporocysts. Dogs fed infected heart muscle stored at -4 degrees C for 48 h did not shed sporocysts, but those fed similar infected tissues stored at -2 degrees C for 24 h shed sporocysts. The results indicate that sarcocysts of S. levinei are rendered noninfective by heating to 65 degrees C or by freezing at -4 degrees C.     

Effect of temperature during induced moulting on plumage renewal and subsequent production

1. Four groups of 18 cages each containing 4 brown laying hybrids aged 79 weeks were formed by two levels of restriction of a moulting diet (lasting for 29 d) being applied at two moulting temperatures (11 degrees C and 29 degrees C, lasting for 56 d). 2. Low temperature-low feeding resulted in greatly retarded growth of remiges, but the final extent was similar in all 4 groups, and reached 4 to 5 new primaries (median value). Body-plumage of hens moulted at 11 degrees C was 25% heavier than of hens moulted at 29 degrees C. 3. Second year production variables (rate of laying, egg mass, efficiency of food utilisation) were greatly influenced by moulting temperature (low moulting temperature performing better), but not by feeding rate. 4. The persistence of the improved food utilisation is related to energetic consequences of improved plumage renewal during moulting at the lower temperature, which can be seen as an acclimatization effect.     

Purification and some properties of a Bacillus cereus mouse lethal toxin.

A mouse lethal toxin (MLT) produced by Bacillus cereus isolated in vomiting-type food poisoning was purified by chromatography on DEAE-Sephadex A-25 followed by gel filtration on Sephadex G-75. Purified MLT possessed a molecular weight of 33,000-34,000. It showed mouse lethality and hemolytic (HL) activity on sheep and rabbit erythrocytes; the latter erythrocytes were more weakly hemolyzed than the former ones. However, fluid accumulation in mouse ligated intestinal loops was not induced by purified MLT at the highest concentration used. Both MLT and HL activities were stable at pH 6-9, during storage at -20 degrees C for 8 weeks, and resistant to papain, cholesterol, lecithin, and dithiothreitol treatments. Most activity was lost during storage at 4 degrees C or 25 degrees C for 2 weeks or upon treatment with trypsin, trypanblue, or ethanol. The activities were resistant to heating at 37 degrees C for 5 min, less resistant at 98 degrees C for 5 min, and sensitive at 60 degrees C for 5 min. It can be concluded from the results that MLT is different from the diarrheagenic toxin produced by B. cereus isolated in diarrheal-type food poisoning, but is similar to, if not identical, hemolysin II.     

Effects of air velocity on convective and radiant heat transfer from domestic fowls at environmental temperatures of 20 degrees and 30 degrees C

The effects of air movement upon sensible heat loss from individual birds at ambient temperatures of 20 degrees and 30 degrees C were determined by partitional calorimetry using a newly developed wind tunnel calorimeter. The relationship between area specific convective heat loss (W/m2) and air velocity (in the range 0.3 to 1.05 m/s) was described by y = 56.5 + 16.9 log x at an air temperature of 20 degrees C, but at 30 degrees C convective heat loss increased linearly with air speed (y = 11.8 + 40.1 x). At 20 degrees C sensible heat exchange (W/m2) was related to air velocity by y = 70.6 exp (0.099 x) and by y = 41.4 + 25.9 x at 30 degrees C, the proportional increase with air speed being greater at the higher temperature. The differences in the effects of air movement on convective cooling and sensible heat loss at 20 degrees and 30 degrees C reflect the thermoregulatory responses of the birds, induced by the thermal demands of the environment.