Survival of larvae of Boophilus annulatus, Boophilus microplus, and Boophilus hybrids (Acari: Ixodidae) in different temperature and humidity regimes in the laboratory.

The survival period for larvae of Boophilus annulatus (Say), Boophilus microplus (Canestrini) and hybridized Boophilus ticks was determined by exposure to various combinations of temperature (20, 25, 30 and 35°C) and relative humidity (32, 63, 75, 84 and 97% RH) in the laboratory. Results indicated that within a given temperature and RH regime, there was no difference (P> 0.05) in larval survival among the three species tested, indicating that these ticks respond similarly over a wide range of temperature and RH combinations. Larval survival in all three species was longest (P < 0.05) at 20°C and either 84 or 97% RH. With each increase in temperature at the 84 and 97% RH treatment levels, there was a corresponding significant (P < 0.05) decrease in larval survival. When the temperature reached 35°C at all humidities or when the RH was 63% or less at all temperatures, the mean larval survival period was 43 days or less in all cases and little difference (P> 0.05) was observed among the treatment regimes included. Results suggest that at a RH of 75% and more, the temperature is the determining factor in larval survival, whereas at a RH of 63% and less the RH is the determining factor in larval survival, regardless of temperature.     

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.     

Hepatozoon canis infecting dogs in the State of Espírito Santo, southeastern Brazil

From May 2007 to March 2008, blood samples were collected from 92 healthy dogs living in 21 households (17 farms in rural area, and 4 homes in urban area) in 6 counties of the State of Espírito Santo, southeastern Brazil. In addition, ticks were collected from these dogs. A mean of 4.4 ± 3.0 dogs (range: 1–12) were sampled per household; 78 and 14 dogs were from rural and urban areas, respectively. Polymerase chain reaction (PCR) designed to amplify fragments of the 18S rDNA gene of Babesia spp or Hepatozoon spp revealed amplicons of the expected size in 20 (21.7%) dogs for Babesia, and 54 (58.7%) dogs for Hepatozoon. All Babesia-positive dogs were also Hepatozoon-positive. Among the 21 households, 15 (71.4%) from 3 counties had at least one PCR-positive dog, including 13 farms (rural area) and 2 homes (urban area). A total of 40 PCR products from the Hepatozoon-PCR, and 19 products from the Babesia-PCR were submitted to DNA sequencing. All generated sequences from Hepatozoon-PCR were identical to each other, and to corresponding 18S rDNA sequences of H. canis in GenBank. Surprisingly, all generated sequences from the Babesia PCR were also identical to corresponding 18S rDNA sequences of H. canis in GenBank. Dogs from 10 rural and 2 urban households were found infested by Rhipicephalus sanguineus ticks. Immature of Amblyomma cajennense ticks were found in dogs from only 4 rural households (also infested by R. sanguineus). All but one household with R. sanguineus-infested dogs had at least one Hepatozoon-infected dog. Statistical analysis showed that the presence of ticks (i.e. R. sanguineus) infesting dogs in the households was significantly (P < 0.05) associated with at least one PCR-positive dog. There was no significant association (P > 0.05) between PCR-positive dogs and urban or rural households. Canine hepatozoonosis caused by H. canis is a high frequent infection in Espírito Santo, Brazil, where it is possibly vectored by R. sanguineus. Since all infected dogs were found apparently healthy, the pathogenicity of H. canis for dogs in Espírito Santo is yet to be elucidated.     

Neem-tree (Azadirachta indica Juss.) extract as a feed additive against the American dog tick (Dermacentor variabilis) in sheep (Ovis aries)

Acaricides can be conveyed to ticks via the blood of their hosts. As fruit and kernel extracts from the Meliaceae family, and, in particular the tetranortriterpenoid azadirachtin (AZA) inhibits tick egg production and embryogenesis in the Ixodidae ticks, we investigated the effects of Neem Azal^®, an extract containing 43% AZA, given as a feed additive to lambs artificially infested with engorging adult Dermacentor vairiabilis ticks. After tick attachment, the lambs were allotted to three dietary treatments: AZA0 (control, n = 10), AZA0.3 (n = 5), and AZA0.6 (n = 5), with feed containing 0%, 0.3%, and 0.6% AZA on DM basis, respectively. In half of the AZA0 lambs, ticks were sprayed on day 4 after attachment with an ethanol:water:soap emulsion containing 0.6% AZA (AZA0S). In spite of its very pungent odor, the neem extract was well accepted by all but one lamb. No differences were found between treatment groups in liver enzymes in blood, and there was no indication of toxicity. The plasma AZA concentrations after 7 and 14 days of feeding AZA were (4.81 and 4.35 μg/mL) for the AZA0.6 and (3.32 and 1.88 μg/mL) for the AZA0.3 treatments, respectively (P < 0.0001). Treatments were not lethal to ticks, but tick weights at detachment were 0.64, 0.56, 0.48, and 0.37 g for ticks from the AZA0, AZA0.3, AZA0S, and AZA0.6 treatments (P < 0.04), respectively, suggesting that blood AZA impaired blood-feeding. The highest mortality rate after detachment was for AZA0.6 (P < 0.09). As AZA affects embryo development and ticks at the molting stages, we expect that following treatments of hosts for longer periods, one-host ticks will be more affected than the three-host tick D. variabilis.     

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.     

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 high environmental temperatures on physiological and nutritional status of prepubertal Holstein heifers

A series of studies were carried out to determine the effects of increasing environmental temperature (20 to 28 to 33 °C) at a constant (60%) relative humidity on the physiological responses, energy and nitrogen balance of prepubertal Holstein heifers (n = 4, initial body weight = 198 ± 3 kg). The relatively high environmental temperatures of 28 and 33 °C increased rectal temperature by 0.2 and 1.2 °C and increased respiration by 23 and 58 breaths per min, respectively. At 28 °C, dry matter intake was similar to that at 20 °C, whereas at 33 °C dry matter intake was lower (P < 0.10) by about 9%. However, the dry matter digestibility was higher (P < 0.05) at 33 °C than at 20 °C, hence the thermal treatments did not affect daily weight gain. At 33 °C blood urea nitrogen increased whereas hematocrit and plasma glucose concentrations decreased. At 33 °C energy and nitrogen intake were lower (P < 0.10), while energy and nitrogen digestibility were higher at 33 °C than at 20 °C. Urinary nitrogen was elevated at 33 °C resulting in lower nitrogen retention. Although, energy retention as fat did not differ (P > 0.10) among the treatments, energy retention as protein was lower (P < 0.05) at 33 °C than at 20 °C or 28 °C. These results confirmed that the effects of environmental temperature on prepubertal heifers were severe above 28 °C.     

Effect of δ-aminolevulinic acid on growth performance, nutrient digestibility, blood parameters and the immune response of weanling pigs challenged with Escherichia coli lipopolysaccharide

This study investigated the effects of dietary δ-aminolevulinic acid (ALA) on growth performance, nutrient digestibility, blood parameters and whether ALA improved the immune response of weanling pigs challenged with Escherichia coli lipopolysaccharide (LPS). Eighty pigs (body weight = 7.21 ± 0.51 kg) were allotted to four dietary treatments, with four pens per treatment and five pigs per pen. Basal diets were supplemented with 0, 5, 10, and 15 mg/kg ALA (as-fed basis) and fed for 35 days. At the end of the feeding period, 10 pigs were selected from both the 0- and 10-mg/kg ALA treatment groups; five were injected i.p. with LPS (50 μg/kg BW) and the other five pigs with an equivalent amount of sterile saline, resulting a 2 × 2 factorial arrangement. Blood sample and rectal temperature data were collected at 0, 2, 4 and 12 h after challenge. Growth performance was not affected by dietary treatments over the total experimental period. However, dry matter (DM) and nitrogen (N) digestibility was improved in the 15-mg/kg ALA treatment group at day 35 (P < 0.05). Serum hemoglobin (Hb) and iron levels were also increased, with the 10-mg/kg ALA treatment showing the highest concentration (P < 0.05). On day 35, red (RBC) and white blood cell (WBC) counts were elevated, with the 5- and 10-mg/kg ALA treatments having the highest counts (P < 0.05). During challenge, LPS injection elevated rectal temperature at 2 and 4 h postchallenge (P < 0.05). Plasma cortisol concentration was also increased by LPS injection at 2 and 4 h postchallenge and an ALA-alleviating effect was evident at 2 h postchallenge (P < 0.01). Concentration of plasma insulin-like growth factor-I (IGF-I) was increased in the ALA-supplemented treatments at 2 h postchallenge (P < 0.05). LPS injection increased plasma tumor necrosis factor-α (TNF-α) concentrations at 2, 4 and 12 h (P < 0.01), while an ALA-alleviating effect was observed at 2 and 4 h postchallenge (P < 0.05 and P < 0.10, respectively). Challenge with LPS decreased WBC counts at 2 and 4 h postchallenge (P < 0.01). At 12 h postchallenge, RBC, WBC and lymphocyte counts were affected by LPS challenge, while an ALA effect was only observed on WBC count (P < 0.05). In conclusion, dietary supplementation of ALA in weanling pigs can improve DM and N digestibilities, and iron status and have a beneficial effect on the immune response during inflammatory challenge.     

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.     

Comparison of Caudal Epidural Anesthesia with Lidocaine-distilled Water and Lidocaine-MgSO4 Mixture in Horses

This study was performed to compare the onset and duration of analgesia produced by either a lidocaine−MgSO₄ or lidocaine−distilled water combination administration in the caudal epidural space of horse. Seven healthy adult horses, aged 11.7 ± 1.4 years (mean ± SD), body weight (kg) 567 ± 32.5 (mean ± SD), were selected for this study. Caudal epidural anesthesia was produced in all horses by administering 2% lidocaine (0.22 mg/kg) diluted in 1 mL distilled water and repeated with 2% lidocaine (0.22 mg/kg) diluted in 1 mL 10% MgSO₄ 2 weeks later. Time to onset (minutes), duration (minutes), and cranial spread of epidural analgesia were recorded. Heart rate (HR), respiratory rate (RR), and body temperature (°C) were recorded. Measurements were taken at 0 (as a baseline value before epidural administrations) and at 5, 10, 15, 30, 60, and 75 minutes after the epidural administrations of each treatment. Statistical analyses included paired Student t test and analysis of variance (computer program SPSS, Analytical Software, version 15.00). Statistical significance was set at P < .05. Onset of analgesia was significantly different (P < .001) between lidocaine-distilled water (2.38 ± 0.47 minutes) and lidocaine−MgSO₄ (4.62 ± 0.54 minutes). Duration of analgesia after lidocaine−MgSO₄ (186.0 ± 7.0 minutes) was longer than lidocaine-distilled water (54.5 ± 7.3 minutes). No significant differences were recorded for HR, RR, and body temperature in comparison with baseline values for each group. Using the lidocaine−MgSO₄ combination for obstetric and surgical procedures could commence relatively soon after epidural injection and could be completed without readministration of anesthetic agent.     

Molecular screening and risk factors of enterotoxigenic Escherichia coli and Salmonella spp. in diarrheic neonatal calves in Egypt

The aim of the present study was to carry out molecular epidemiological investigation on enterotoxigenic Escherichia coli (ETEC) K99 and Salmonella spp. in diarrheic neonatal calves. Fecal samples were obtained from 220 diarrheic calves at 9 farms related to four governorates in central and northern Egypt. E. coli and Salmonella spp. isolates were examined for E. coli K99 and Salmonella spp. using PCR. ETEC K99 was recovered from 20 (10.36 %) out of 193 isolates, whereas Salmonella spp. was recovered from nine calves (4.09%).Multivariable logistic regression was used to evaluate the risk factors associated with both infections. ETEC K99 was significantly affected by age (P < 0.01; OR: 1.812; CI 95%: 0.566–1.769), colostrum feeding practice (P < 0.01; OR: 5.525; CI 95%: 2.025–15.076), rotavirus infection (P < 0.001; OR: 2.220; CI 95%: 0.273–1.251), vaccination of pregnant dams with combined vaccine against rotavirus, coronavirus and E. coli (K99) (P < 0.001; OR: 4.753; CI 95%: 2.124–10.641), and vitamin E and selenium administration to the pregnant dam (P < 0.01; OR: 3.933; CI 95%: 0.703–1.248).Infection with Salmonella spp. was found to be significantly affected by the animal age (P < 0.05; OR: 0.376; CI 95%: 0.511–1.369), Hygiene (P < 0.05; OR: 0.628; CI 95%: 1.729–5.612), and region (P < 0. 01; OR: 0.970; CI 95%: 0.841–1.624).The results of the present study indicate the importance of PCR as rapid, effective and reliable tool for screening of ETEC and Salmonella spp. when confronted with cases of undifferentiated calf diarrhea. Moreover, identification of the risk factors associated with the spreading of bacteria causing diarrhea may be helpful for construction of suitable methods for prevention and control.     

Energy balance, milk production and reproduction in grazing crossbred cows in the tropics with and without cereal supplementation

The aim of this work was to study the effect of parity order and reproductive management systems on rabbit production performance. A total of 73 rabbit does (I group) were submitted to a 35-day intensive rhythm [artificial insemination (AI) on day 4 post-partum (pp) and weaning at 25 days of lactation], and 108 rabbit does (SI group) were submitted to a 42-day semi-intensive rhythm (AI on day 11 pp and weaning at 35 days of lactation) during 9 months. Primiparous does had the lowest live body weight at parturition (P < 0.0001) and at 21 days of lactation (P < 0.0001). They also had lower milk production (P < 0.0001) than does with later parities. I group animals needed a higher number of AI than SI group to become pregnant (1.70 ± 0.03 vs. 1.39 ± 0.03; P < 0.0001: especially after the third). Prolificacy was not affected by the management system. Parturition interval (PI) was longer than expected in both groups [56.0 ± 1.4 and 50.9 ± 1.38 days in I and SI groups, respectively (P < 0.05)]. Mean productivity, estimated as number of weaned rabbits per female and year, was 12 kits higher in rabbit does of the SI group (P < 0.05). From the third parturition onward, an increase in live body weight of kits at different ages was observed. At 21 (P < 0.05) and 25 days of age (P < 0.01), kits from the I group rabbit does weighed more than those from the SI group; however, the latter showed a higher weight at 35 (P < 0.05) and 60 days of age (P < 0.05). Rabbit does with two or three parturitions had higher litter size at 21 and 25 days of age (P < 0.0001 and P < 0.001, respectively). Kit mortality between 21 and 25 days of age and between 35 and 60 days of age was not affected by treatments but was higher in the I group between 25 and 35 days (18.2 vs. 5.03% in the I and SI groups, respectively; P < 0.0001) and as age of does increased (P < 0.05). In light of these results, we could conclude that long term doe reproductive performance is negatively affected and litter viability decreased when using intensive compared to a semi-intensive reproductive management.     

Dietary supplementation with Acanthopanax senticosus extract enhances gut health in weanling piglets

The present study was conducted to investigate the effects of Acanthopanax senticosus extract (ASE) as a dietary additive on gut health in weanling piglets by examining diarrhea frequency, intestinal microbiota and morphology. A total of 96 Duroc× (Landrace × Yorkshire) piglets weaned at 21 days of age with an average initial body weight (BW) of 5.6 ± 0.4 kg were randomly assigned to 3 treatment groups with 4 duplicates of 8 piglets each. The piglets were fed basal diet to which had been added 0 or 1 g/kg of ASE, or 0.7 g/kg antibiotics, respectively. Fecal consistence was monitored twice daily and the frequency of diarrhea was calculated. On day 21 after the initiation of supplementation, 8 piglets were randomly selected from each treatment group (2 piglets per pen) and slaughtered. The jejunum, ileum, colon and cecum were then excised and fixed in 10% neutral formalin solution to determine villus height and crypt depth, after their contents were collected to determine microbiota. The results showed that dietary supplementation with ASE increased (P < 0.05) the density of bacterial populations that co-migrated with Lactobacillus amylovorus, Lactobacillus salivarius, Bacillus subtilis, and Clostridium lituseburens, but decreased (P < 0.05) those co-migrating with Staphylococcus aureus, Salmonella typhimurium, Ruminococcus forques, and E. coli O157:H7 in the PCR-DGGE profiling analysis when compared with the control group. The villus height of the duodenum, jejunum and ileum increased (P < 0.05) by 14.8, 13.7 and 10.0%, while the crypt depth decreased (P < 0.05) by 17.9, 9.1 and 12.1%, respectively, in response to dietary ASE supplementation. Additionally dietary supplementation with ASE or an antibiotic decreased (P < 0.05) the frequency of diarrhea by 55.6 and 52.2%, respectively, compared with the control group. In conclusion, these findings suggest that dietary supplementation with ASE could regulate the microbiota composition and maintain a normal morphology of gut mucosa in weanling piglets, thereby decreasing diarrhea that resulted from weaning stress.     

Effects of hypotaurine, cysteamine and aminoacids solution on post-thaw microscopic and oxidative stress parameters of Angora goat semen

This study was conducted to determine the effects of cysteamine, hypotaurine and aminoacids solution (BME) on standard semen parameters, lipid peroxidation and antioxidant activities of Angora goat semen after the freeze–thawing process. Ejaculates collected from four Angora goats were evaluated and pooled at 37 °C. Semen samples, which were diluted with a Tris-based extender containing the antioxidants hypotaurine (5 mM) and cysteamine (5 mM), and an aminoacid solution (13%), and an extender containing no antioxidants (control), were cooled to 5 °C and frozen in 0.25-ml French straws in liquid nitrogen. Frozen straws were thawed individually at 37 °C for 20 s in a water bath for evaluation.Supplementation with cysteamine, hypotaurine and BME caused significant (P < 0.05) increases in sperm motility, and significant (P < 0.05) decreases in total abnormality rates in comparison to the control group. While all in vitro treatments did not affect the acrosomal abnormality rates, hypotaurine and BME but not cysteamine significantly (P < 0.05) increased the HOST results as compared to the control group. Supplementation with antioxidants and BME did not significantly affect MDA levels and CAT activity in comparison to the control group (P > 0.05). The antioxidants hypotaurine and cysteamine decreased SOD activity when compared to the BME group and controls (P < 0.001).     

Serum profile of metabolites and hormones in obese (Iberian) and lean (Landrace) growing gilts fed balanced or lysine deficient diets

Serum hormones and metabolites of lean and obese gilts were determined. Fifteen Iberian and fifteen Landrace gilts, of approximately 20 kg body weight, were fed isoenergetic (13.6 kJ ME/g DM, semisynthetic diets either with equilibrated amino acid profile at two crude protein (CP) levels (12% (0.81% lysine (Lys)) and 16% CP (1.08% Lys) as-fed basis) or lysine deficient (12% (0.30% Lys) or 16% CP (0.36% Lys) as-fed basis for Iberian and Landrace, respectively). Lysine deficient diets were offered only at the optimal protein level for maximum protein accretion for each breed. Each dietary treatment was assayed in 5 animals. Gilts were allocated in metabolic cages at 21 ± 1.5 °C with free access to water and fed four times daily at 90% ad libitum during 10 days. On day 11th blood samples were taken 5–5.5 h postprandial and serum obtained and frozen at − 20 °C. Growth hormone (GH), insulin like growth factor I (IGF-1), insulin, leptin, glucose, urea, creatinine, cholesterol and triglycerides were determined. Landrace gilts had higher serum glucose (P < 0.05) and creatinine (P < 0.001) than Iberian while no differences (P = 0.122–0.494) were found for the rest of the biochemical variables. Higher serum levels of insulin, IGF-1 and leptin (60, P < 0.05; 79, P < 0.05 and 189%, P < 0.001, respectively) and no differences in GH (P = 0.512) were encountered in Iberian gilts compared to Landrace. Dietary crude protein did not alter the serum hormonal profile (P = 0.110–0.454). Serum cholesterol (total, P < 0.01; HDL, P < 0.05; LDL, P < 0.05) decreased and triglycerides and urea increased (P < 0.05 and P < 0.001, respectively) as dietary crude protein increased. When lysine deficient diets were used, serum glucose decreased (P < 0.05) and urea increased (P < 0.001) compared to equilibrated diets. Serum insulin (P = 0.052), IGF-1 (P < 0.05) and leptin (P = 0.051) concentration decreased when lysine deficient diets were fed to the animals while GH remained unaffected (P = 0.214).These data suggest that Iberian and Landrace growing gilts have distinct hormone and metabolite serum profiles which are altered by a severe dietary lysine deficiency.     

Mixed-breed twin production by contralateral direct-transfer of male embryos to beef cow recipients bred to GnRH-based, fixed-time AI

Two experiments were conducted to develop protocols for the use of fixed-time artificial insemination and embryo transfer (TAI and TET, respectively) to increase beef cattle productivity. Suckled beef cows were given GnRH (100 µg im) on Day − 10, and PGF (25 mg im) on Day − 3, with TAI on Day 0 (66 h later), and assigned to either embryo recipient (ER) or no embryo (NR) treatments on Days 6 or 7. Semen from Gelbvieh (GB) beef sires was used for TAI; sexed-male in vivo developed Holstein embryos (HO) were placed nonsurgically (TET) into the uterine horn contralateral to the corpus luteum. In Experiment 1, ovarian status of cows (n = 111; 69 ± 11 d postpartum; mean ± SD on Day 0) in Groups I and II was presynchronized with a single PGF treatment on Day − 24; Groups II and III received GnRH concurrent with TAI, and ER (n = 78) were selected from all groups on Days 6 or 7. Neither presynchronization nor GnRH affected rates of recipient selection, Day 45 pregnancy (43.2, 43.2, and 54.0% for Groups I, II and III, respectively), or calving (40.5, 37.8, and 43.2%). However, treatment with GnRH increased HO-birth rate (8.0, 14.0 and 24.0%; P < 0.05). In Experiment 2, cows (n = 99, 113 ± 10 d postpartum) were assigned on Day 7 to NR and ER, with and without hCG (2500 IU im), following a TAI protocol (as per Group III in Experiment 1). Treatment with hCG increased reproductive rate (1.16 vs 1.44 calves/calving; P < 0.05), but had no significant effect on rates of Day 45 pregnancy (45.3 vs 41.3%), calving (31.2 vs 37.0%) or HO-births (20.0 vs 26.0%). In summary, GnRH-based synchronization rates were 71.2% (based on circulating progesterone concentrations that were < 1.0 ng/mL on Day 0 and ≥ 1.0 ng/mL on Day 7); TET did not affect GB-birth rate, but more calves were produced by ER than NR cows (1.43 vs 1.02 calves/calving; P < 0.01); and weaned calf production was 53% greater for twin- than singleton-suckled cows (392 ± 25 vs 256 ± 11 kg/dam; mean ± SEM, P < 0.05). Therefore, GnRH-based TAI and TET protocols for mixed-breed twin production increased beef cow productivity.     

Reference values on hematologic parameters of the Brazilian donkey (Equus asinus) breed

Thirteen hematologic measurements were performed on 38 healthy donkeys of the Brazilian breed (32 females and 6 males) aged 3 to 19 years. Mean ±SD (minimum-maximum) values were as follows: red blood cells (RBC) 6.82 ± 0.67 (5.46−8.17) × 10⁶/μL; packed cell volume (PCV) 37.63 ± 2.76 (32.00−44.00) %; hemoglobin (Hb) 12.87 ± 0.98 (10.70−14.50) g/dL; mean corpuscular volume (MCV) 55.45 ± 4.06 (48.90−62.70) fL; mean corpuscular hemoglobin (MCH) 18.85 ± 1.14 (17.40−21.60) pg; mean corpuscular hemoglobin concentration 34.20 ± 1.24 (31.50−36.40) g/dL; white blood cells (WBC) 8.22 ± 1.49 (4.60−11.50) × 10³/μL; bands 1.77 ± 2.00 (0.00−9.00) %; segmented neutrophils 41.18 ± 5.98 (30.50−53.20) %; lymphocytes 50.53 ± 6.25 (35.30−63.50) %; monocytes 1.57 ± 1.30 (0.00−4.60) %; eosinophils, 4.64 ± 2.17 (1.00−10.00) %; basophils 0.33 ± 0.58 (0.00−2.90) %. It was demonstrated that values of some blood parameters of the Brazilian donkey breed were similar to results obtained for other donkey breeds. The results of this study serve as reference ranges for donkey populations and can be useful for health control, which in turn helps in the diagnosis and prognosis of diseases.

Introduction

Blood examination has been performed for several reasons: as a screening procedure to assess general health; as an adjunct to patient's infection; and to evaluate the progress of certain disease states.[1] However, there are important differences in physiology, behavior, and management that influence the common diseases encountered in donkeys. Successful diagnosis and treatment often depend on recognizing these differences. [2] Unfortunately, a limited number of observations have been reported on the hematologic values for donkey breeds and populations and these studies are far from complete. [3, 4, 5, 6, 7, 8 and 9] Hematologic variation values for donkey breeds and populations should be associated with the age, sex, and the time of sampling in relation to exercise, geographic, and nutritional factors. Perhaps some of the variations observed among different authors were also associated with different techniques and methods. [4]Apart from the investigations by Perdigão de Oliveira et al,[3] Gacek et al, [10] and Mori et al, [11] we could find no published record of blood values of the Brazilian donkey breed. The aim of this investigation was to determine reference ranges for hematologic analyses in the Brazilian donkey breed to establish a baseline for further scientific and clinical use.

Materials and methods

The material for the present study consisted of 38 adult donkeys of the Brazilian breed (32 females and 6 males), ranging between 3 and 19 years of age (mean ±SD: 8.3 ± 3.5 years), from the Estação Experimental de Colina do Instituto de Zootecnia da Secretaria de Agricultura do Estado de São Paulo, and kept under similar feeding and management conditions. All animals were apparently healthy, and they were handled carefully to reduce any possible effects of stress on the parameters analyzed.Blood samples were draw from the jugular vein into 5 mL evacuated glass tubes containing EDTA as anticoagulant. Blood smears were prepared immediately after being collected from whole blood and then were air-dried. The collected blood samples were immediately stored at 4°C and processed within 6 hours.RBC and WBC counts were performed using an electronic counter (Serono Baker). Hb concentration was determined by the cyanmethemoglobin method. PCV was determined after the blood had been transferred to microhematocrit tubes and centrifuged at 10,000 g for 5 minutes. MCV, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration were calculated from PCV, Hb concentration, and RBC count.[1] Blood smears were stained with modified May-Grunwald Giemsa stain for determination of the differential WBC count. At least 200 WBCs were counted for differential WBC determinations.All analyses were performed within the Departamento de Clinica Médica da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo.Data analysis was performed using a statistical software (Graph Pad InStat version 3.01, 32 bit for Windows 95/NT). Each parameter was tested for normality applying the Kolmogorov and Smirnov methods. The reference ranges for the hematologic analyses were given as mean ±SD and the interval from minimum and maximum values.Unpaired t-tests were used to determine the effect of sex (female versus male) on hematologic values, for data normally distributed. If data did not seem to be normally distributed, non-parametric tests (Mann-Whitney U test) were used. The level of statistical significance was set at P < .05.

Results

Reference ranges of hematologic constituents of the blood of the Brazilian donkey breed determined in the current study are shown in Table 1 and Table 2. Most of the values had Gaussian distribution (P < .10). Only basophils did not seem to be normally distributed (P < .05). Comparisons of hematologic ranges of the Brazilian donkeys with other donkey breeds and populations are shown in Table 1 and Table 2.     

Growth and carcass characteristics of lambs in relation to plasma IGF-I and some histological traits of Longissimus lumbarum and Biceps femoris as affected by breed and age at slaughter

Ten male lambs of each of Rahmani (R) and Ossimi (O) were used to study the growth and carcass characteristics within their first year of age (360 days). After suckling period (4 months), lambs were fed on the regular ration. During the course of the experiment, body weight (BW) was recorded monthly accompanied by blood sampling to determine IGF-I. Four lambs from each breed were slaughtered at 270 and 360 days of age, and Longissimus lumbarum (L. lumbarum) at the 9th, 10th and 11th ribs and Biceps femoris (B. femoris) muscles were separated to be used for chemical analysis and histological study.Breed had no effect on the growth curve of the two studied breeds. Chemical analysis indicated that (O) had higher fat (0.9%, P < 0.0001) and lower (0.3%, P < 0.01) moisture compared to (R), with no difference in protein and ash contents. The concentration of IGF-I was almost similar in both breeds up to day 270 of age, and then it was higher in (R). Number of fibers per bundle was higher (P < 0.05) in (O) than in (R) by 6.1%, while, the other histological traits were similar in the two studied breeds.Chemical analysis indicated that lambs slaughtered at 360 days of age had lower (0.7%, P < 0.0006) moisture and higher fat (17.3%, P < 0.0001) compared to day 270. Up to day 270 of age, IGF-I concentration was < 500 ng/ml before increasing to > 500 ng/ml up to the end of the experiment. No effect of age was observed on the studied histological traits.L. lumbarum muscle contained higher protein (4.2%, P < 0.0001), fat (104%, P < 0.0001) and ash (10%, P < 0.001), while moisture was less (5.6%, P < 0.0001) compared to B. femoris muscle. B. femoris muscle was higher (P < 0.0001) in number and diameter of fibers, bundle and stroma sectional areas relative to L. lumbarum by 38.2, 78.8, 53.2 and 23.8%, respectively.     

The use of a semiochemical bait to enhance exposure of Amblyomma variegatum (Acari: Ixodidae) to Metarhizium anisopliae (Ascomycota: Hypocreales)

Experiments were conducted to explore the use of a semiochemical bait to enhance exposure of Amblyomma variegatum Fabricius (Acari: Ixodidae) to different formulations of the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorok. (Ascomycota: Hypocreales). Initially, the relative efficacies of attraction-aggregation-attachment pheromone (AAAP), made up of o-nitrophenol, methyl salicylate and nonanoic acid in the ratio 2:1:8, 1-octen-3-ol and butyric acid, were evaluated in an olfactometer. Only AAAP and 1-octen-3-ol were found to elicit attractive responses to the tick. Simultaneous release of 1-octen-3-ol and AAAP together with CO₂ from a trap in semifield plots attracted up to 94.0 ± 6% of adult ticks from a distance of 6 m, and up to 24.0 ± 5.1% from 8 m. Formulations of M. anisopliae (dry powder, oil, and emulsifiable) applied within the trap baited with AAAP, 1-octen-3-ol and CO₂ resulted in high levels of contamination of the ticks attracted to the traps. However, 48 h after autoinoculation, 89.1 and 33.3% of conidia were lost in dry powder and oil formulations, respectively. Emulsifiable formulation showed least loss of propagules (17.1%). Samples of ticks attracted to the baited traps were transferred to plastic basins containing grass and maintained for 5 weeks. The experiment was conducted in rainy and dry seasons. Emulsifiable formulation gave the highest relative tick reduction in both seasons: 54.7 and 46.5% in rainy and dry seasons, respectively, followed by oil formulation (32.0 and 23.8%) and powder formulation (38.0 and 24.4%).