Effect of blood sample handling post-collection on Erysipelothrix rhusiopathiae antibody titres

A study was conducted to determine the effect of blood sample mishandling on the performance of an enzyme-linked immunosorbent assay for the detection of antibodies against Erysipelothrix rhusiopathiae. Eleven sample maltreatments (storage at −10 °C, storage at 4 °C, heat treatment of clotted blood, haemolysis, repetitive freeze–thaw cycling, and substitution of plasma in place of serum) were simulated in a laboratory environment and then run concurrently against a gold standard sample (storage at −80 °C).The mishandling treatment groups that simulated high levels of haemolysis had significantly lower optical density (OD) readings when compared to the gold standard. However, the magnitude of the effects was relatively small and only samples with OD values close to the cut-off changed state from positive to negative. Heat treatment had a minor, but non-significant, effect on OD values. Findings from this study suggested that immunoglobulin G antibody was stable in the face of most common sample mishandling events.     

Life-history parameters of Culicoides (Avaritia) imicola Kieffer in the laboratory at different rearing temperatures

This laboratory study investigates the sub-adult developmental cycle of field collected Culicoides (Avaritia) imicola Kieffer (Diptera; Ceratopogonidae). The period required from blood-feeding field-collected females to the production of progeny adults occupied 34–56 days at 20 °C, 15–21 days at 25 °C and 11–16 days at 28 °C, demonstrating clear temperature dependence. When reared at 28 °C, C. imicola demonstrated higher variability in fecundity (between 2.4 and 20.6 eggs/female) and lower hatching rates (50.0–62.2%), although larval survival rates to pupation were low at all temperatures (20–30%). Similarly, the mean emergence rate from pupae was the highest at lower temperatures. These results highlight the difficulty in establishing and maintaining a laboratory colony of this species from field-collected material and results are discussed in reference to future research directions that may aid this process.     

Effects of intermittent suckling on the performance and digestive efficiency of Iberian piglets weaned at 35 days of age

During two consecutive replicates Iberian sows (n = 20) were subjected either to a conventional lactation (C) or intermittent suckling (IS). Piglets in the C treatment had free access to the dam. Piglets on IS treatment were separated from the sow during 6 h on days 29 and 30, 8 h on days 31 and 32, and 10 h on days 33 and 34. Litters on both treatments were weaned at 35 days of age and offered a starter diet until day 60 of age. The apparent digestibility (ApD) of nutrients and gross energy (GE) of the post-weaning diet was measured using Cr₂O₃ as external indigestible marker. Litters on IS tended to increase solid feed intake during the milk restriction (125 ± 12 vs. 70 ± 19 g piglet^− 1 day^− 1; P = 0.08). Feed intake was improved by IS during the second week post-weaning (676 ± 17 vs. 497 ± 12 g piglet^− 1 day^− 1, P < 0.01), but differences were totally offset after the 7th week of age. Differences in average growth rate of litters on C and IS treatments did not attain statistical significance during the period of restriction and in the first week post-weaning. Litters on IS showed increased growth rate during the second week post-weaning (422 ± 31 vs. 289 ± 33 g piglet^− 1 day^− 1; P = 0.01), but slower daily gains from the 3rd week post-weaning to 60 day of age than litters on C treatment (371 ± 14 vs. 432 ± 15 g piglet^− 1 day^− 1; P < 0.05). Nevertheless, neither at weaning nor at 60 days of age average body weight of piglets differed between treatments (7.71 ± 0.31 vs. 7.94 ± 0.30 kg for C and IS, at 35 days of age, 15.62 ± 0.65 vs. 16.20 ± 0.62 kg, at day 60). The apparent digestibility of nutrients of the starter diet offered after weaning remained unaffected by the treatment, except for a trend for higher ApD of GE for the piglets on IS treatment (P = 0.08). ApD for CP and GE was 78.1 ± 0.6 and 78.8 ± 0.3%, respectively. No significant differences in the proportions of total viscera and gastrointestinal tract to empty body weight (EBW) were observed between C and IS piglets at weaning and 60 d of age. In conclusion, the increase in feed intake observed prior and early after weaning as a consequence of intermittent suckling during the last week of 35 d lactation had no effect on nutrient digestibility measured at two weeks after weaning and failed on improving Iberian piglet performance.     

Resistance of Santa Ines and crossbred ewes to naturally acquired gastrointestinal nematode infections

This trial was carried out in Piracicaba, São Paulo State, Brazil, to comparatively evaluate the degree of resistance to naturally acquired gastrointestinal nematode infections in sheep of the following genetic groups: purebred Santa Ines (SI), SI crossbred with Dorper (DO × SI), Ile de France (IF × SI), Suffolk (SU × SI), and Texel (TE × SI). Fifteen ewes from each group were raised indoors until 12 months of age. At this age, they were moved to pasture that was naturally contaminated by nematode infective larvae and were evaluated from December to May, 2007. Rainfall ranged from 267 mm in January to 37 mm in April. Maximum and minimum mean temperatures ranged from 32.5 °C to 19.0 °C in March and from 25.9 °C to 12.8 °C in May. There was an increase in the mean number of eggs per gram of feces (EPG) after animals were placed on pasture with significant difference between the SI (80 EPG) and IF × SI (347 EPG) groups in January; and the DO × SI (386 EPG) and TE × SI (258 EPG) groups in May. The highest mean fecal egg count (FEC), 2073 EPG, was recorded for the TE × SI group in February. All groups showed a progressive reduction in body weight throughout the experiment of 12.0% (TE × SI) to 15.9% (SU × SI). In general, the animals with the highest FEC presented the lowest packed cell volumes (PCV); the highest correlation coefficient between FEC × PCV occurred in the SU × SI sheep in January (r = −0.70; P < 0.01). Similarly, there was an inverse relationship between FEC and blood eosinophil values, with the highest correlation coefficient in the TE × SI sheep in February (r = −0.64; P < 0.05). Immunoglobulin G (IgG) levels against Haemonchus contortus antigens increased in all groups as a result of the exposure to parasites and remained relatively constant until the end of the study, with the exceptions of SU × SI and TE × SI, which showed a rise in IgG levels during the last sampling that coincided with a reduction in mean FEC. In conclusion, crossbreeding Santa Ines sheep with any of the breeds evaluated can result in a production increase and the maintenance of a satisfactory degree of infection resistance, especially against H. contortus and Trichostrongylus colubriformis, the major nematodes detected in this flock.     

The effect of high calcium intake by pregnant Awassi ewes at late gestation on minerals status and performance of ewes and newborn lambs

Calcium (Ca) requirement increase rapidly by pregnant ewes at late gestation as a result of high fetal growth. Furthermore, Ca deficiency results in poor growth and bone development in growing newborn lambs. The aim of this study was to evaluate the effect of calcium supplementation (Calcium carbonate–CC) and vitamin AD₃E injection (75,000 IU/shot) to pregnant Awassi ewes at late gestation (− 45 days prepartum) on their mineral levels and performance of their newborn lambs. Forty pregnant Awassi ewes at late gestation (65 ± 6.1 kg) were selected before parturition (− 45 days; using hormonal sponges for estrus synchronization) and randomly divided to 4 dietary treatments. The treatments were: group 1 Control (1.4% CC); group 2 (2.4% CC); group 3 (1.4% CC + 2 shots vitamin AD3E at − 45 and − 21 days prepartum); group 4 (2.4% CC + 2 shots vitamin AD3E at − 45 and − 21 days prepartum). Blood samples were collected from ewes (− 45, 0 days prepartum: 30 and 60 days postpartum) and lambs (0, 30 and 60 days old). Average milk yield and samples were collected. Body weights of lambs were taken at birth, 30 and 60 days old. Blood serum and milk samples were prepared and analyzed by AAS for Ca, Mg, Co, Mn and Cu concentrations.Calcium concentrations in bloods serum of ewes, 0 and 60 days postpartum, and lambs at weaning (60 days old) from group 4 were significantly (P < 0.05) higher compared to other groups. No significant effect (P > 0.05) of treatment on Co, Mg and Cu in blood serum of ewes and lambs except for Cu concentration in ewes. Moreover, a significant increase (P < 0.01) in milk inorganic% (30 and 60 days samples) in ewes from group 2 when compared with other groups. The average milk yield (AMY) of ewes from group 2 and 4 at late stage of lactation (45–60 days) were significantly higher (P < 0.05) compared to groups 1 and 3. In addition, Ca% in colostrums samples from ewes in groups 2 and 4 were significantly higher (P < 0.01). The average daily gain (ADG) and total body gain for lambs up to weaning (60 days old) were significantly higher (P < 0.05) in groups 1 and 2 compared with the other groups.In conclusion, increasing level of Ca intake in feed of Awassi ewes at late gestation, with or without vitamin AD₃E injections, improve the calcium concentration in colostrums and blood serum of ewes and their lambs without negative effect in their health and performance.     

Pharmacokinetics of marbofloxacin after a single oral dose to loggerhead sea turtles (Caretta caretta)

The single-dose disposition kinetics of marbofloxacin (MBX) were determined in clinically healthy loggerhead sea turtles (n = 5) after oral (PO) administration of 2 mg kg⁻¹ bodyweight. Marbofloxacin plasma concentrations were determined by DAD–HPLC (LOD/LOQ 0.015/0.05 μg ml⁻¹). Data were subjected to non-compartmental analysis. Following PO administration, marbofloxacin achieved maximum plasma concentrations of 11.66 ± 2.53 mg L⁻¹ at 15.00 ± 3.00 h. The absence of general adverse reactions in the turtles of the study, and the favourable pharmacokinetic properties (long half-life and high maximum plasma concentration) of MBX administered PO at the single-dose of 2 mg kg⁻¹ suggest the possibility of its safe and effective clinical use in loggerhead sea turtles.     

Correlation between clinical signs of depth of anaesthesia and cerebral state index responses in dogs during induction of anaesthesia with propofol

The cerebral state index (CSI) is used for monitoring EEG and depth of anaesthesia. The objective of this study was to analyse the correlation between ocular reflexes, CSI and estimated propofol plasma concentrations (PropCP) in dogs during induction of anaesthesia with propofol.Fourteen dogs were premedicated with acepromazine 0.05 mg kg⁻¹ IM. Anaesthesia was induced with a 200 ml h⁻¹ propofol 1% constant infusion rate until loss of corneal reflex using RugLoop II software with Beths’ pharmacokinetic model to estimate PropCp.Palpebral reflex (PR) and the corneal reflex (CR) were tested every 30 s and classified as present (+) or absent (−), and eyeball position was registered as rotated ventromedialy (ERV) or centred (EC).Heart rate (HR), mean arterial pressure (MAP) and CSI values were analyzed from baseline before the beginning of propofol infusion (T0) until loss of CR; CSI and PropCp, CSI and anaesthetic planes, and PropCp and anaesthetic planes were compared using correlation analysis.PropCp reached 7.65 ± 2.1 μg ml⁻¹ at the end of the study. CSI values at T0 were 89.2 ± 3.8. Based on the observation of ocular reflexes and eyeball position, it was possible to define five anaesthetic planes: A (superficial) to E (deep), being A (PR+/CR+/EC), B (PR+/ERV/CR+), C (PR−/ERV/CR+), D (PR−/EC/CR+) and E (PR−/EC/CR−). There was a significant correlation between PropCp and the anaesthetic planes (R = 0,861; P < 0.01). No significant correlation was observed between CSI and the anaesthetic planes or between CSI and PropCp. MAP decreased significantly from T0 until loss of corneal reflex (from 98 ± 14 mmHg to 82 ± 12 mmHg); HR did not change significantly (from 101 ± 30 bpm to 113 ± 16 bpm).The CSI monitoring was not consistent with the clinical observations observed in the different stages of depth anaesthesia. This could limit the use of CSI for monitoring depth of anaesthesia with propofol.     

Antibody-ELISA for Trypanosoma evansi: Application in a serological survey of dairy cattle, Thailand, and validation of a locally produced antigen

Trypanosoma evansi is generally considered a mild pathogen in bovines. However, in Asia, acute and chronic signs have been observed in cattle, with high levels of parasitaemia, abortion and death. Investigations in Asian cattle are needed to better understand this epidemiological situation. To generate comparable data at a regional level, development and standardization of an antibody-enzyme linked immunosorbent assay for T. evansi (ELISA/T. evansi) was initiated and applied in an epidemiological survey carried out in dairy cattle in Thailand. A batch of 1979 samples was collected from dairy farms located throughout the country's four regions. Soluble T. evansi antigens initially produced in France were also produced in Thailand for comparison and technology transfer. Screening of 500 samples allowed us to identify reference samples and to determine the cut-off value of the ELISA. Seropositive animals – some of them confirmed by PCR – were found in the four regions, in 12 out of 13 provinces, in 22 out of 31 districts, in 56 farms out of 222 (25%, 95%CI ± 6%) and in 163 animals out of 1979 (8.2, 95%CI ± 1.2%). Estimated seroprevalence in 35 farms ranged between 1% and 30%, and in 21 farms it was >30%. Approximately 25% of survey cattle were exposed to the infection, in various situations. A sub-sample of 160 sera was tested on both antigens. Wilcoxon's (Z = 1.24; p = 0.22) and McNemars's tests (CHI2 = 3.55; p = 0.09) did not show any significant differences, showing that the locally produced antigen is suitable for further evaluation in the surrounding countries. Use of this standardized serological method will broaden knowledge of the prevalence and impact of the disease at the regional level in South-East Asia. Further validation of this ELISA will be necessary in other host species such as buffalo, horse and pig.     

Protection against Toxoplasma gondii brain cyst formation in mice immunized with Toxoplasma gondii cytoskeleton proteins and Lactobacillus casei as adjuvant

The aim of the study was to evaluate the protection generated in mice against Toxoplasma gondii brain cyst burden by vaccination with T. gondii cytoskeleton proteins using Lactobacillus casei as adjuvant. One hundred and sixty-eight NIH mice were randomly allocated into eight groups of 21 mice each. Animals were immunized as follows: in group 1 with Toxoplasma lysate antigen (TLA) in Freund's modified adjuvant, containing L. casei (FMA), in group 2 with Toxoplasma cytoskeleton proteins (TCPs) in FMA, in group 3 with FMA, in group 4 with phosphate buffered saline (PBS), in group 5 with L. casei dead by heath (Lc), in group 6 with Freund's complete adjuvant (FCA), in group 7 with TLA in FCA, and in group 8 with TCP in FCA. Mean brain cyst burden (±S.E.M.) was assessed in mice 8 weeks after challenge with T. gondii Me49 strain (20 cysts per mouse). The percentages of reduction in cyst burden per brain (P < 0.01) as compared with the group 4 (control: mean 3181 ± 97.5) were 77.25% (724 ± 98) in group 1, 88.02% (381 ± 97.5) in group 2, 38.92% (1943 ± 130.3) in group 3, 44.31% (1771.4 ± 102) in group 5, 59.28% (1295.2 ± 99.1) in group 7 and 55.69% (1409.5 ± 89.9) in group 8. In order of importance, the best protection was obtained in groups 2, 1, 7, 8, 5 and 3. Noticeably the mice inoculated with L. casei alone showed a significant reduction in T. gondii brain cysts (P < 0.01), while those animals treated with FCA alone did not. Additionally, IgM anti-T. gondii antibody levels, as determined by ELISA 2 weeks after challenge, were highest in group 2 (P < 0.01) than in the other seven groups. Results suggest that T. gondii cytoskeleton proteins with L. casei as adjuvant constitute a good anti-toxoplasmosis vaccine candidate.     

Effects of Saccharomyces cerevisiae on ruminal pH and microbial fermentation in dairy cows: Yeast supplementation on rumen fermentation

An experiment was conducted with eight ruminally-cannulated cows using a crossover design with 2 periods to determine the effects of yeast supplementation on rumen fermentation. Holstein dairy cows in late lactation were either supplemented with 0.5 g/hd/d of Saccharomyces cerevisiae, an active dry yeast (CNCM-1077, Levucell SC20 (r) SC, Lallemand Animal Nutrition) or not supplemented (control). A basal diet consisting of 60% forage and 40% concentrate (DM basis) was fed once daily to both groups of cows throughout the entire experiment. Ruminal pH was measured continuously every 22 min using a pH probe that was placed in the ventral rumen sac for 6 consecutive days. Volatile fatty acid and ammonia N concentrations in the rumen were measured on days 5 or 6 of the 12-d period for each cow and DM intake was monitored throughout the experiment. Data were analyzed using a mixed-effects model with repeated measures. There were no differences in dry matter intake between treatments. Mean ruminal pH was greater (P < 0.05) when yeast was supplemented (6.53 ± 0.07) compared with the control (6.32 ± 0.07). Average maximum and minimum ruminal pH were also greater (P < 0.05) when yeast was supplemented (7.01 ± 0.09 and 5.97 ± 0.08, respectively) compared with the control (6.80 ± 0.09 and 5.69 ± 0.09, respectively). Time spent under the subacute acidosis threshold, pH less than 5.6, was lower (P < 0.05) with yeast supplementation compared with control cows. No difference was observed for ruminal ammonia N concentrations (mean = 14.0 ± 1.2 mg/dL) between treatments. Total VFA concentration (mM) in the rumen tended to be lower (P = 0.10) in the yeast-supplemented cows (107.3 ± 6.35) than in the control cows (122.4 ± 6.35), which could be related to the greater pH observed with yeast supplementation. Supplementing dairy cows with active dry yeast in the current experiment increased the mean, minimum and maximum ruminal pH; decreased time spent in subacute rumen acidosis, and tended to decrease total VFA concentration in the rumen compared with control cows.     

Responses to condensed tannins of flowering sulla (Hedysarum coronarium L.) grazed by dairy sheep: Part 1: Effects on feeding behaviour, intake, diet digestibility and performance

The concentration of condensed tannins (CT) in sulla (Hedysarum coronarium L.) is moderate and overall regarded as beneficial. However, the intake of this forage can reduce diet digestibility, particularly during flowering phase. An experiment was run to assess the effect of CT on feeding behaviour, intake, diet digestibility and performance of dairy sheep rotationally grazing sulla at flowering phase. Twenty-four late-lactating sheep were blocked in two homogeneous groups and submitted to the following treatments: i) twice daily drenching with 200 g/day of a 50% w/v water solution of an anti-tannic substance, polyethylene glycol, group PEG; ii) twice daily drenching with 200 g/day of water, group CON (Control group). All the sheep rotationally grazed as a flock two sulla plots from April to June (8 weeks in total). Sward height, herbage mass, botanical and chemical composition of the herbage were measured at the beginning and the end of each grazing period. The feeding behaviour (3 sheep per group) was continuously monitored for 24 h in 6 weeks using the IGER behaviour recorders. Herbage DM intake (DMI), dietary DM digestibility (DMD) and apparent CP digestibility (CPD) were estimated on 8 sheep per group by the n-alkane method. On average, PEG group had longer total grazing (503 ± 12 vs 460 ± 12 min, P < 0.05) and eating time (425 ± 13 vs 391 ± 13 min, P < 0.07) than CON group. Moreover PEG group showed shorter inter-meal intervals (41 ± 3 vs 52 ± 3, min, P < 0.05) and higher number of daily meals than CT-exposed group (24 ± 1 vs 19 ± 1 min, P < 0.01). The herbage DMI was not affected by the treatment whereas DMD (74.60 ± 3.48 vs 58.30 ± 3.01%), and CPD (60.14 ± 4.83 vs 38.21 ± 4.83%) were both increased by PEG administration (P < 0.05) confirming the negative effect of sulla CT on these variables. Milk yield tended to be higher in PEG than CON (1331 ± 45 vs 1205 ± 59 ml, P < 0.11). Milk protein content was similar between groups while milk fat content was higher in CON than PEG ewes (6.61 ± 0.15 vs 6.11 ± 0.15%, P < 0.05), being the reverse true for milk urea (46.04 ± 1.27 vs 53.04 ± 0.76%, P < 0.001). To conclude, this experiment shows that when sulla is grazed at flowering as monoculture, dietary CT can exert negative effects on DM and CP apparent digestibility, in this study partially counterbalanced by a better metabolic utilisation of the nutrients up-taken.     

Influence of methionine and dithioerythritol on sperm motility, lipid peroxidation and antioxidant capacities during liquid storage of ram semen

The aim of this study was to investigate the effects of methionine and dithioerythritol, added to the Tris extender, on ram sperm motility and LPO (lipid peroxidation) and antioxidant capacities during liquid storage up to 72 h at 5 °C. Ejaculates collected from five Merino rams, were evaluated and pooled at 37 °C. This study included two experiments. In experiment 1, each pooled ejaculate was divided into four equal aliquots and diluted (37 °C) with the base extender, containing 0 (control), 1, 2 and 4 mM methionine, at a final concentration of approximately 4 × 10⁸ sperms/ml (single step dilution), in a 15-ml plastic centrifuge tube. In experiment 2, dithioerythritol, at concentrations of 0 (control), 0.5, 1 and 2 mM, was used as an additive in the extender, and the procedure explained above was applied for the division of aliquots and the dilution of semen. Diluted semen samples were kept in glass tubes and cooled from 37 to 5 °C in a cold cabinet, and maintained at 5 °C. Sperm motility and LPO and total glutathione (GSH) and glutathione peroxidase (GPx) capacities were determined at 5 °C for periods of 0, 24, 48 and 72 h of liquid storage.The extender supplemented with 1 mM methionine led to higher motility percentages (77.0 ± 1.2%), in comparison to the control group (66.0 ± 4.9%), during 72 h of liquid storage (P < 0.05). As regards dithioerythritol, it did not statistically improve the motility rates for any of the storage times at 5 °C. In biochemical assays, differences in LPO levels between the groups with antioxidants and the control groups were not statistically significant. Compared to the control group, no significant difference was observed in GSH and GPx activities following the addition of methionine, during 72 h of storage. Total GSH and GPx activities did not increase significantly upon supplementation with 0.5 and 1 mM of dithioerythritol, compared to the control group, at any of the time points (P > 0.05). Dithioerythritol at 2 mM led (P < 0.01) to elevating GSH activity, compared to the control group, during 72 h of liquid storage. GPx activity was approximately 10 times higher for 2 mM of dithioerythritol (P < 0.001), compared to that of the control group at all time points.The question regarding the sustainability of sperm survival, LPO and antioxidant capacities following liquid storage of semen remains unanswered. Further studies are required for a better understanding of the biochemical changes and to obtain more information on the determination of lipid peroxidation and antioxidant capacities during cooled storage of ram semen.     

Effects of GnRH in combination with PGF2α on the dynamics of follicular and luteal cells in post-pubertal Holstein heifers

Holstein heifers were randomly allotted by weight, age and body condition score to one of three treatments to test the hypothesis that GnRH administration concurrent with PGF_2α injection would advance follicle or corpus luteum (CL) development parallel to an induced luteolysis of the pre-existing CL. Heifers in the control group (n = 14) received two treatments of PGF_2α(25 mg, im) given 10 days apart. Groups 2 (n = 14) and 3 (n = 14) received an additional treatment of GnRH (100 μg, im) after the first and second PGF_2α respectively. Estrus detection began immediately after PGF_2α and continued for 80 h. Blood sampling was initiated 7 days prior to the first PGF_2α (day − 7) and continued on days 0, 7, 10 (prior to the second PGF_2α), 17 and 24. Heifers were artificially inseminated after the second PGF_2α and pregnancy diagnosed at 60 days. There was a trend (P < .10) toward a lower estrus response in group 3 when compared to the other groups. Pregnant heifers in group 2 had lower progesterone (0.44 ± 0.09 vs. 1.72 ± 0.56 ng/ml) a week after the second PGF_2α than the non-pregnant animals in that group (P < .05). Similar results were observed in the control group but only within the responding heifers (0.61 ± 0.08 vs. 0.93 ± 0.03 ng/ml; P < .05). Progesterone in heifers in group 2 remained high on day 0, 7, and 10 (1.48 ± 0.37, 1.23 ± 0.39, 1.96 ± 0.36 ng/ml) in spite of the treatment with PGF_2α. This data suggest that administration of GnRH following PGF_2α alters bovine luteal and/or follicular cell function.     

Development and evaluation of an indirect enzyme-linked immunosorbent assay for the detection of antibodies against Campylobacter fetus in cattle

Campylobacteriosis is a zoonosis that occurs worldwide. Infection with Campylobacter fetus (C. fetus) causes infertility and abortion in sheep and cattle. The current study focuses on the SapA gene of C. fetus that encodes surface array proteins and plays an important role in the virulence of C. fetus. The SapA-N (1398 bp) and SapA-C (1422 bp) fragments were amplified from the C. fetusSapA gene using polymerase chain reaction (PCR), and the corresponding recombinant proteins rSapA-N and rSapA-C were expressed in Escherichia. coli BL21 cells. Results of Western blotting and enzyme-linked immunosorbent assay (ELISA) showed that the immunological activity of rSapA-N was higher than that of rSapA-C (P < 0.05). Therefore, rSapA-N was selected to establish an indirect ELISA for detecting antibodies against C. fetus. The diagnostic criteria were as follows: S/P ? 0.45: positive; S/P < 0.4: negative; 0.45 > S/P ? 0.4: suspected. The specificity and sensitivity of our method were 94.3% and 88.6%, respectively. Moreover, no cross-reactions were observed between rSapA-N and serum samples that were positive for other bovine bacterial pathogens diseases such as Mycobacterium avium subspecies paratuberculosis. One hundred and two serum samples from cows that had experienced abortion were tested. Four and 2 C. fetus-positive serum samples were found among the 70 bovine brucellosis-positive samples and the 32 infectious bovine rhinotracheitis (IBR)-positive samples, respectively. The findings suggest that the rSapA-N-based ELISA method has immense potential in future applications.     

Effects of transportation during the hot season and low voltage electrical stimulation on histochemical and meat quality characteristics of sheep longissimus muscle

The effect of transportation during the hot season (42 °C) and low voltage electrical stimulation on physiological, histochemical and meat quality characteristics of Omani sheep was studied. Forty intact male sheep were divided into two equal groups: 3 h transported or non-transported. The non-transported group remained in holding pens for 48 h prior to slaughter, while the transported group was transported 300 km (approximately 3 h) in an open truck under solar radiation on the day of slaughter. Blood samples were collected from the animals before loading and prior to slaughter. Fifty percent of the carcasses from each group were randomly assigned to low voltage (90 V) at 20 min postmortem. Temperature and pH decline of the left longissimus thoracis muscle were monitored. Ultimate pH, WB-shear force, sarcomere length, myofibrillar fragmentation index (MFI), expressed juice, cooking loss and color L, a, b were measured on samples from both sides muscles collected at 24 h postmortem at 3–4 °C. The transported sheep had significantly higher plasma cortisol (P < 0.01), adrenaline, nor-adrenaline and dopamine concentrations (P < 0.05) than non-stimulated animals. Electrical stimulation resulted in a significantly (P < 0.05) more rapid pH fall in muscle during the first 12 h after slaughter. Muscles from electrically-stimulated carcasses had significantly (P < 0.05) lower pH values, longer sarcomere length, lower shear force value, higher expressed juice, MFI and lighter L than those from non-stimulated ones. The muscle samples from the transported sheep had significantly (P < 0.05) smaller and lower proportion of Types I and IIA fibers than those from the non-transported group. This study indicated that pre-slaughter transport at high ambient temperatures can cause noticeable changes in physiological and muscle metabolism responses in sheep. Electrical stimulation improved meat quality characteristics, which indicate that meat quality of transported sheep can be improved by electrical stimulation.     

Pathogenic 'Bison-type' Mycobacterium avium subspecies paratuberculosis genotype characterized from riverine buffalo (Bubalus bubalis) in North India

Despite low per-animal productivity of ruminants in developing countries, Johne's disease has not been investigated in buffaloes, which are primarily found in these countries. This is due to lack of expertise, diagnostic kits and priority to production diseases like Johne's disease. Presence of pathogenic Mycobacterium avium subspecies paratuberculosis (Map) was investigated by screening of target tissues (mesenteric lymph nodes and large intestine) by culture and IS 900 PCR, in 50 sacrificed buffaloes. Indigenous ELISA kit originally developed for goats and sheep was standardized in buffaloes and used to estimate sero-presence of Map in 167 serum samples representing population of buffaloes in Agra region of North India. In culture, 48.0% buffaloes were positive from 50 tissues each from mesenteric lymph nodes (34.0%) and large intestine (36.0%). IS 900 PCR was standardized using specific primers (150 C and 921) and 229 bp-amplified product was characteristic for Map. Of the 25 mesenteric lymph nodes, 40.0% were positive in IS 900 PCR. Genomic DNA from Map cultures was successfully amplified from all the 24 isolates (100.0%). Map was further genotyped as 'Bison type' using IS 1311 PCR-REA. Culture of tissues showed high presence of Map in target tissues, despite high culling rate in buffalos in view of high demand of buffalo meat. Specific tissue-PCR provided rapid confirmation of Map infection in sacrificed buffaloes. In tissue-PCR, all the cultures were positive as compared to 40.0% detected directly from tissues. ELISA kit using indigenous protoplasmic antigen was highly sensitive as compared to commercial antigen in detecting Map infection therefore, could be used as 'Herd Screening Test' in buffaloes against Johne's disease. This pilot study first time reports a highly pathogenic 'Bison-type' genotype of M. avium subspecies paratuberculosis from the riverine buffaloes (Bubalus bubalis) of Agra region in North India.     

Lincomycin and spectinomycin in the treatment of breeding rams with semen contaminated with ureaplasmas

Ureaplasma species were isolated from semen samples collected sequentially from one Awassi and three Assaf breeding rams. Each ram was injected subcutaneously with an aqueous solution of lincomycin and spectinomycin for five consecutive days at a dose equivalent to 4·5 mg kg⁻¹ lincomycin and 9·0 mg kg⁻¹ spectinomycin daily. Serum and semen samples were collected at intervals during the treatment and assayed for lincomycin. No Ureaplasma species were isolated from semen samples collected during the course of the treatment and at intervals for 17 days after the last treatment. The concentration of lincomycin in semen ranged from 0·51 μg ml⁻¹ four hours after treatment to 0·08 μg ml⁻¹ 24 hours after treatment, and these levels were three to nine times higher than the corresponding serum concentrations.     

Prevalence of Michigan dairy herds infected with Mycobacterium avium subspecies paratuberculosis as determined by environmental sampling

A cross-sectional, stratified random survey of Michigan dairy herds was conducted to estimate the prevalence of herds infected with Mycobacterium avium paratuberculosis (MAP), the causative agent of Johne's disease, in Michigan using targeted environmental sampling. One pooled sample each from the primary manure storage area and a high-traffic common cow area from each herd was collected and cultured for MAP using the ESP^® culture system II. A herd was classified as positive if at least one sample was culture positive for MAP. State, agricultural district, and herd size stratum prevalence were calculated. Information on past MAP testing and cattle purchase history was collected, and logistic regression was performed to determine their importance to the MAP status of the herd. One hundred twenty-seven herds were contacted, and 94 agreed to participate in the study. The environment of 38 (40.4%) herds cultured positive for MAP. MAP was found in all herds (n = 15) with greater than 200 lactating cows. Herds that had tested for MAP or purchased cattle in the previous 5 years were 4.6 and 3.1 times, respectively, more likely to be infected than herds that had not. MAP continues to be prevalent on Michigan dairy farms, especially those with greater than 200 lactating cows. The environmental sampling protocol used in this study is an economically attractive alternative for monitoring herd level prevalence and the progress of Johne's disease control programs at the state or national level. Implementation of such a program would aid states in monitoring Johne's control program progress, and guide changes over time.     

Influences of dietary regimens on microbial content in gastrointestinal tracts of meat goats

This experiment was conducted to evaluate the effects of dietary treatments on microbial loads and pH of gastrointestinal tract contents in meat goats, as well as the concentration of volatile fatty acid (VFA) in the rumen. Crossbred (Boer x Spanish) goats (n = 36; BW = 17.7 kg) were assigned randomly to one of three experimental diets (n = 12/diet or 3 pens/treatment) for 90 days:alfalfa (Medicago sativa) hay alone (AH-diet); 18% CP concentrate alone (C-diet); or, a combined diet (AHC-diet), consisting of the AH-diet for the first 45 days, followed by 45 days of the C-diet. After evisceration, pH values of rumen liquor and colon digesta were immediately measured from each animal, as well as aseptically collected rumen liquor and rectal samples to determine the microbial loads. Collected rumen liquor was also prepared for volatile fatty acid (VFA) contents. Feeding meat goats with alfalfa hay alone had higher (P < 0.05) rumen (7.17) and colon (7.10) pH compared with those fed either the concentrate alone or combined-diet. Although the acetate content was high in the AH-fed group (66.3 mM) compared to the AHC-diet group (34.6 mM), no significant differences were found in the total VFA contents in rumen liquor among the goats fed three different dietary regimens. Total plate counts were not significantly different among goats fed the experimental diets in the rumen or rectal samples. Escherichia coli counts in the rectal samples were lower (P < 0.05) in the AH-diet group (6.43 log₁₀ CFU/g) compared with the C-diet (8.21 log₁₀ CFU/g) or AHC-diet (8.40 log₁₀ CFU/g) groups. However, no significant differences were found in the E. coli counts of rumen samples from goats fed the experimental diets. The mean (± SEM) rumen E. coli counts were 1.38, 1.65, and 2.51 ± 0.560 log₁₀ CFU/g in the AH-, C-, and AHC-diet groups, respectively. The results indicate that feeding hay alone may decrease the fecal shedding of E. coli in meat goats with increasing the rumen and colon pH.     

Rumen degradation characteristics of sweet potato foliage and performance by local and crossbred calves fed milk and foliage from three cultivars

The study investigated rumen dry matter (DM) degradability characteristics in a completely randomized design and the effects of milk, sweet potato foliage (SPF) from three cultivars (A = TIS-87/0087; B = TIS-8164; C = TIS-2532.OP.1.13), dried brewers' grains (DBG) and cottonseed meal (CSM) as supplements to Panicum maximum (Panicum) for pre-weaned calves in randomized complete block designs. Diet 1 = milk + SPF-A foliage + Panicum, Diet 2 = milk + SPF-B foliage + Panicum, Diet 3 = milk + SPF-C foliage + Panicum, and Diet 4 = milk + DBG & CSM + Panicum (as control). Dry matter (130 ± 0.4 to 864 ± 3.9 g kg^− 1), ash (54 ± 4.2 to 173 ± 2.8 g kg^− 1 DM), OM (827 ± 4.2 to 946 ± 5.7 g kg^− 1 DM), N (7.4 ± 0.6 to 38.6 ± 1.4 g kg^− 1 DM), and NDF (439 ± 1.4 to 774 ± 8.5 g kg^− 1 DM) contents were highly significant (P < 0.01). In Trial I, 16 pre-weaned calves were used over 70 d with milk intake (34.8 ± 4.4 ml kg W^− 0.75 d^− 1), Panicum DMI (22.3 ± 2.77 g kg W^− 0.75 d^− 1), total DMI (35.7 ± 2.83 g kg W^− 0.75 d^− 1), and LWG (198 ± 44.6 g d^− 1) not significantly different (P > 0.05). Supplement DMI varied (P < 0.05) from 11.6 g kg W^− 0.75 d^− 1 in Diet 3 to 16.6 g kg W^− 0.75 d^− 1 in Diet 4. In Trial II, 16 pre-weaned local and crossbred calves were involved over 77 d with initial age of calves, Panicum intake, metabolic DMI, and LWG similar (P > 0.05) among crosses. Birthweight varied (P < 0.05) from 17.3 kg for N'Dama × Jersey crosses to 21.2 kg for White Fulani × Brown Swiss crosses. Supplement and total DMI ranged (P < 0.05) from 172 to 483 g d^− 1 for N'Dama × Jersey crosses to 233 and 674 g d^− 1 for non-inseminate or purebred calves, respectively. The LWG in the White Fulani × Brown Swiss and the N'Dama × Jersey calves were respectively 30% and 24% better, though not significantly, than purebred calves. In Trial III, rumen DM degradability characteristics of feeds in three N'Dama steers showed no significant differences (P > 0.05) in slowly degradable fraction (b) and rate of degradation of b (c). Soluble fraction (a), 48-h degradation, potential degradability (PD) and effective degradability (ED) varied significantly (P < 0.05) and were lowest in Panicum, but similar for foliage among the three sweet potato cultivars. Panicum fodder showed improvements in degradation characteristics with supplementation.