Pilot study--parenteral treatment of recently acquired subclinical mastitis during lactation

OBJECTIVE: To conduct a pilot study to investigate the effect of parenteral antibiotic treatment with penethamate hydriodide on recently acquired sub-clinical mastitis. DESIGN: Sixty-nine cows from five Western Victorian dairy farms were enrolled in a pilot trial. Cows that had an Individual Cow Cell Count (ICCC) of greater than 250,000 cells per mL in their milk at their first or second herd test were enrolled in the trial if they were in their first lactation, or if they had a history of ICCCs below 250,000 cells per mL at all herd tests during the previous lactation. These criteria were used in an effort to exclude cows with chronic subclinical mastitis and include cows with recently acquired subclinical mastitis. PROCEDURE: Cows were divided into control and treatment groups on the basis of having an odd or even ear tag number. The treatment group was treated with penethamate hydriodide shortly after the first or second herd test; the control group was left untreated. The ICCCs of subsequent herd tests were recorded for the remainder of lactation. RESULTS: In 58% of the treatment group, and 25% of the control group, all subsequent ICCCs for that lactation were less than 250,000 cells per mL (p < 0.05). If only first and second lactation cows were considered, 72% of the treatment group, and 11% of the control group, had subsequent ICCCs less than 250,000 cells per mL (p < 0.05). CONCLUSION: An ICCC of greater than 250,000 cells per mL is a good predictor for more high ICCCs at subsequent herd tests. This effect was significantly altered in our trial by treatment with penethamate hydriodide at or near the first or second herd test. Whilst in general treatment of high ICCC cows during lactation is considered to be unrewarding, this pilot study suggests that treatment of cows and heifers appropriately selected on the basis of a previous history of low ICCCs can lead to sustained reductions in cell counts. These results support the case for further study into this area.     

Optimization of conditions for in vitro production of radical oxygen species and expression of tissue factor by canine mononuclear cells and granulocytes for use in high-throughput assays

The purpose of this study was to optimize conditions for high throughput measurement of radical oxygen species (ROS) production and expression of tissue factor, also termed procoagulant activity, by canine leukocytes. Granulocytes and mononuclear cells were separated by density gradient centrifugation from peripheral blood collected on several occasions from three healthy large breed dogs. To determine optimal conditions for ROS production, granulocytes were incubated for 1 or 3h in PBG (PBS containing 0.5% BSA and 5mM glucose) or RPMI containing 10% fetal bovine serum (FBS); lipopolysaccharide (LPS), zymosan, peptidoglycan (PGN) and phorbol myristate acetate (PMA) were used as stimuli. ROS was assessed by conversion of the nonfluorescent dye dihydrorhodamine 123 to fluorescent rhodamine 123 by radical species released into the media. To identify optimal conditions for expression of tissue factor, mononuclear cells were incubated for 5h in RPMI containing different concentrations of heat-inactivated FBS (HI-FBS), and LPS, zymosan, PGN or PMA as stimuli. Expression of tissue factor was determined using a one-stage recalcification assay performed in an automated nephelometric coagulation analyzer. Neither LPS nor zymosan increased ROS production by granulocytes incubated in PBG media. In contrast, granulocytes incubated in RPMI had dose-dependent increases in ROS production in response to zymosan and PGN. ROS production was significantly increased by incubation with concentrations of LPS of 0.01microg/ml or greater, and by zymosan concentrations of 0.1microg/ml or greater. ROS production in response to incubation with PMA was significantly increased starting at 10(-7)M, and was significantly greater for cells incubated in RPMI than cells incubated in PBG. LPS-, zymosan- and PGN-stimulated procoagulant activity increased in a dose-dependent manner, whereas PMA-stimulated procoagulant activity peaked at 10(-7)M. Increasing concentrations of HI-FBS significantly increased LPS-, zymosan- and PGN-induced procoagulant activity of mononuclear cells. Results obtained in this study indicate production of ROS by canine granulocytes is optimal when these cells are incubated for 3h in RPMI with LPS (0.1microg/ml), zymosan (10 microg/ml), PGN (10 microg/ml), and PMA (10(-7)M). Furthermore, canine mononuclear cells express procoagulant activity in response to LPS, zymosan, PGN, and PMA, and responses to LPS, zymosan and PGN are enhanced by the addition of HI-FBS. These findings suggest that HI-FBS retains important serum proteins that facilitate interactions between each of these bacterial or yeast derived products and the mononuclear cells. Consequently, future studies regarding the regulation of procoagulant activity by canine mononuclear cells should be performed in the presence of HI-FBS. Both assays utilized in this study allow high throughput of samples, and therefore are appropriate choices for rapid screening of conditions and/or therapeutic interventions affecting the canine inflammatory system.     

Bovine herpesvirus 4-associated postpartum metritis in a Spanish dairy herd

In more than 10 Spanish dairy cows, a bovine herpesvirus 4 (BHV4) associated postpartum metritis was confirmed by virus isolation, BHV4-glycoprotein B (gB) PCR and/or serology. In this study, 12 cows with, and, at the time of sampling, 3 cows without clinical signs of acute postpartum metritis from one large dairy herd in Spain were examined for bacterial and viral infections. Blood, placenta/caruncles and uterine contents were collected between day 1 and day 20 post-calving, and examined for the presence of bacteria and for viruses by virus isolation, BHV4 DNA by BHV4-gB PCR and/or BHV4 antibody titres. Bovine herpesvirus 4 was detected in 83% of the cases with clinical signs of acute postpartum metritis by virus isolation and/or BHV4-gB PCR. An increase of BHV4 antibodies was detected in all examined postpartum metritis cows and in the 3 cows without clinical metritis. Two of these 3 cows developed severe metritis a few dayss after collecting the first blood sample. A concurrent infections of BHV4 and bacteria, mainly Arcanobacterium pyogenes and Streptococcus sp., were detected in 73% of the examined uterine contents collected from postpartum metritis affected cows. This case-report study showed a clear association between BHV4 infections and acute postpartum metritis in dairy cows. In addition, the BHV4-associated postpartum metritis appeared to be an emerging syndrome in this Spanish herd.     

Frequencies of blood type A, B and AB in non-pedigree domestic cats in Turkey

OBJECTIVES: To determine the distribution of blood types and to estimate the proportion of matings at risk for neonatal isoerythrolysis in non-pedigree domestic cats. METHODS: The present survey determined the frequency of blood types in 301 cats from four distinct regions of Turkey. Ethylenediaminetetraacetic acid-anticoagulated blood samples were typed by simple tube and slide agglutination assays. Serum obtained from type B cats and an anti-B solution, prepared with Triticum vulgaris, were used to determine type A and type B blood, respectively. RESULTS: Of the 301 cats typed, 220 had type A blood, 74 had type B and seven had type AB. There was a significant difference (P<0.01) between the locations of the cats, with fewer type B cats in the eastern than in the western parts of Turkey. Risk for the development of neonatal isoerythrolysis due to A-B mismatch was estimated to be 18.6 per cent. CLINICAL SIGNIFICANCE: The overall type B frequency in Turkish domestic cats is high. Thus, untyped transfusions in these cats carry a high risk of life-threatening acute haemolytic transfusion reactions and neonatal isoerythrolysis. It is therefore strongly recommended that blood typing be performed before breeding or transfusing in order to minimise blood type incompatibility risks.     

Na(+), K(+)-ATPase content in skeletal muscle of dogs with pituitary-dependent hyperadrenocorticism

Several hormones regulate Na⁺, K⁺-ATPase content in the muscle cell membrane, which is essential for maintaining muscle cell excitability. Chronic glucocorticoid excess is associated with muscle weakness and reduced endurance. We hypothesized that chronic glucocorticoid excess affects Na⁺, K⁺-ATPase content in canine skeletal muscle, and contributes to reduced endurance and muscle weakness associated with pituitary-dependent hyperadrenocorticism (PDH) in dogs. Therefore, Na⁺, K⁺-ATPase content in skeletal muscle was evaluated before and after hypophysectomy and hormone replacement (cortisone and l-thyroxin) in dogs with PDH (n = 13), and in healthy controls (n = 6). In addition, baseline and exercise-induced changes in plasma electrolyte concentrations and acid–base balance were evaluated before and after hypophysectomy in dogs with PDH. Na⁺, K⁺-ATPase content of gluteal muscle in dogs with PDH was significantly lower than in control dogs (201 ± 13 pmol/g versus 260 ± 8 pmol/g wet weight; P < 0.01). Similar differences were found in palatine muscle. After hypophysectomy and on hormone replacement, Na⁺, K⁺-ATPase was increased (234 ± 7 pmol/g wet weight). Both plasma pH and base excess in dogs with PDH (7.44 ± 0.01; 1.7 ± 0.6 mmol/l, respectively) were significantly higher (P < 0.05) than after hypophysectomy and hormone replacement (7.41 ± 0.01; −0.2 ± 0.4 mmol/l, respectively). Exercise induced respiratory alkalosis, but did not result in hyperkalemia in dogs with PDH. In conclusion, chronic glucocorticoid excess in dogs with PDH is associated with decreased Na⁺, K⁺-ATPase content in skeletal muscle. This may contribute to reduce endurance in canine PDH, although dogs with PDH did not exhibit exercise-induced hyperkalemia. Na⁺, K⁺-ATPase content normalized to values statistically not different from healthy controls after hypophysectomy and hormone replacement.     

Energy expenditure in Awassi sheep grazing wheat stubble in the northern Negev Desert of Israel

In semi-arid Mediterranean areas, small grain aftermath stubble represents an important summer source of food for grazing flocks of small ruminants. Wheat stubble is a mediocre source of forage and flocks are grazed in summer under harsh conditions of temperature and air dustiness. However, stubble grazing procedures are changing, water and shading are more frequently available between grazing sessions (“improved management”), and the biological soundness of this ancestral practice needs to be re-visited. The present study was aimed at evaluating the cost in energy of “improved” wheat stubble grazing, compared with feeding a similar diet indoors. The intake of stubble was first quantitatively and qualitatively evaluated in Awassi sheep. Ewes consumed daily 980 ± 100 g day^− 1 of wheat stubble. Ewes were then housed and fed diets consisting of wheat hay, straw and grain formulated to be iso-energetic and iso-nitrogenous to diets consumed from wheat stubble. The average intake of ME was similar during the confinement and the pasture periods (6.4 ± 0.5 and 7.6 ± 0.8 MJ day^− 1of ME, respectively). During 2 days of each period, animals were fitted with external electrodes and data loggers of heart rate and skin temperature. Energy expenditure (EE) was calculated from oxygen consumption estimated as the product of heart beats rate measured for the two days by the amount of oxygen delivered to body tissues at each heart beat (O₂ pulse). The O₂ pulse was determined by simultaneously measurement of oxygen consumption and HR twice daily on two occasions, while grazing stubble and indoors. Energy expenditure and energy balance were not different in sheep while grazing wheat stubble (11.1 and − 3.5 MJ day^− 1) or fed indoors (11.1 and − 4.8 MJ day^− 1). Our data show that stubble did not cover nitrogen and energy requirements for maintenance, and that the cost of summer stubble grazing carried out under conditions described here is less than thought before.     

Effects of boar presence on agonistic behavior, shoulder scratches, and stress response of bred sows at mixing

The objective of this study was to determine the effects of boar presence on aggression, shoulder scratches, and salivary cortisol in group-housed sows during the period after mixing, which is when dominance hierarchies are formed. A total of 225 York-shire sows were used. Five groups of 15 sows were each exposed to 1 of 3 levels of boar presence (n = 15): physical (boar in pen with sows; 2.15 m2 per sow), fenceline (boar housed adjacent to sows; 2.3 m2 per sow), or control (no boar in the room; 2.3 m2 per sow). The experiment was divided into 2 phases. In phase 1, behavioral measurements were taken for 48 h after mixing and boar introduction. In phase 2, behavioral measurements were taken beginning on d 6 after mixing and included 24 h before and 48 h after boar removal. In phases 1 and 2, shoulder scratches were scored 24 h before and 24 h after mixing and boar removal, respectively. Saliva samples were collected each morning (0600 to 0700) and afternoon (1600 to 1700) during both phases. Frequencies of intersow aggressive contact (bite, body knock, and head knock) and threats as well as frequencies and durations of fighting bouts were determined from video recordings during daily feeding (49.6 +/- 1.4 min) and nonfeeding (21 +/- 0.2 h) periods. During phase 1, boar presence did not affect the frequency of threats during either the feeding or nonfeeding periods. Control groups had fewer aggressive contacts at feeding during the 25- to 48-h period compared with the fenceline groups (P = 0.01). Total duration of fighting and average fighting bout duration were unaffected by treatment. However, the number of fights occurring during feeding 25 to 48 h postmixing was lower in the physical groups than in the fenceline groups (P = 0.023); measures for the control groups were intermediate. Physical sows also received fewer shoulder scratches postmixing than did control sows (P = 0.048). After mixing, physical sows showed greater morning (P = 0.08) and afternoon (6 h postmixing, P = 0.01; 32 h postmixing, P = 0.08) salivary cortisol concentrations compared with fenceline sows, although they were only numerically greater than those for control sows. During phase 2, removing the boar did not increase fighting among sows or affect shoulder scratches or salivary cortisol concentrations. The presence of a boar was minimally effective at reducing fighting and scratches during the postmixing period, and sows showed a greater stress response in the presence of a boar.     

Environmental factors influencing heat stress in feedlot cattle

Data from 3 summer feedlot studies were utilized to determine the environmental factors that influence heat stress in cattle and also to determine wind speed (WSPD; m.s(-1)) and solar radiation (RAD; W.m(-2)) adjustments to the temperature-humidity index (THI). Visual assessments of heat stress, based on panting scores (0 = no panting to 4 = severe panting), were collected from 1400 to 1700. Mean daily WSPD, black globe temperature at 1500, and minimums for nighttime WSPD, nighttime black globe THI, and daily relative humidity were found to have the greatest influence on panting score from 1400 to 1700 (R2 = 0.61). From hourly values for THI, WSPD, and RAD, panting score was determined to equal -7.563 + (0.121 x THI) - (0.241 x WSPD) + (0.00082 x RAD) (R2 = 0.49). Using the ratio of WSPD to THI and RAD to THI (- 1.992 and 0.0068 for WSPD and RAD, respectively), adjustments to the THI were derived for WSPD and RAD. On the basis of these ratios and the average hourly data for 1400 to 1700, the THI, adjusted for WSPD and RAD, equals [4.51 + THI - (1.992 x WSPD) + (0.0068 x RAD)]. Four separate cattle studies, comparable in size, type of cattle, and number of observations to the 3 original studies, were utilized to evaluate the accuracy of the THI equation adjusted for WSPD and RAD, and the relationship between the adjusted THI and panting score. Mean panting score derived from individual observations of black-hided cattle in these 4 studies were 1.22, 0.94, 1.32, and 2.00 vs. the predicted panting scores of 1.15, 1.17, 1.30, and 1.96, respectively. Correlations between THI and panting score in these studies ranged from r = 0.47 to 0.87. Correlations between the adjusted THI and mean panting score ranged from r = 0.64 to 0.80. These adjustments would be most appropriate to use, within a day, to predict THI during the afternoon hours using hourly data or current conditions. In addition to afternoon conditions, nighttime conditions, including minimum WSPD, minimum black globe THI, and minimum THI, were also found to influence heat stress experienced by cattle. Although knowledge of THI alone is beneficial in determining the potential for heat stress, WSPD and RAD adjustments to the THI more accurately assess animal discomfort.     

Identification of avian strains of Pasteurella multocida in India by conventional and PCR assays

The prevalence of capsular and somatic serotypes were studied among 123 Pasteurella multocida strains isolated from chickens (n = 94), ducks (22), quails (4), turkeys (2) and geese (1) from different geographical regions of India. All strains exhibited similar cultural and morphological characteristics. Ninety-two of the isolates belonged to serotype A:1, the most prevalent serotype, with serotypes A:3, A:1,3, D:3 and F:3 having two isolates each. Only one isolate was positive for serotypes A:4 and D:1. Twenty isolates were untyped. A multiplex capsular PCR assay generated amplicons of sizes 460, 1044, 657 and 854 bp in 106 isolates identified as capsular serotype-A, 15 in serotype D and two in serotype F. Capsular types B and E were not detected in any of the avian isolates studied. The present findings suggest that a multiplex capsular PCR assay may be suitable for the rapid initial identification serotypes P. multocida during epidemiological studies of fowl cholera.