Effects of Bacillus subtilis C‐3102 on production,hatching performance,egg quality,serum antioxidant capacity and immune response of laying breeders

To investigate the supplemental effects of Bacillus subtilis C‐3102 on the production, hatching performance, egg quality, serum antioxidant capacity and immune response of laying breeders, a total of 480 Xuefeng black‐bone (25‐week‐old) hens were randomly assigned into four treatment groups: Hens fed the basal diets with 0 (CON), 3.0 × 10⁵ (BS‐1), 6.0 × 10⁵ cfu/g (BS‐2) and 9.0 × 10⁵ (BS‐3) cfu/g of B. subtilis C‐3102. As the B. subtilis C‐3102 level increased, egg weight (linear, p < 0.01; quadratic, p = 0.003), fertility (linear, p = 0.021; quadratic, p = 0.059), hatchability (linear, p = 0.038; quadratic, p = 0.119) and yolk colour (linear, p = 0.006; quadratic, p = 0.021) increased in a linear or quadratic manner. Yolk index increased quadratically (linear, p = 0.054; quadratic, p = 0.017), and eggshell thickness (linear, p = 0.036; quadratic, p = 0.128), the activity of GSH‐Px (linear, p = 0.024; quadratic, p = 0.078), the concentration of IgM (linear, p = 0.016; quadratic, p = 0.056) and the level of AIV‐Ab (linear, p = 0.034; quadratic, p = 0.103) in the serum increased linearly as dietary supplementation of B. subtilis C‐3102 increased. The results showed that dietary treatments did not affect egg production, feed conversion ratio, egg mass, hatchability of fertile eggs, eggshell‐breaking strength, egg‐shape index, yolk percentage, Haugh unit, T‐SOD, T‐AOC, MDA, IgA and IgG concentrations and the level of NDV‐Ab in the serum. In conclusion, dietary supplementation of 9.0 × 10⁵ cfu/g B. subtilis C‐3102 in laying breeders diets may be a feasible means of effectively increasing egg weight, fertility and hatchability, and improving egg quality such as eggshell thickness, yolk index and yolk colour. Besides, B. subtilis C‐3102 can enhance the activity of GSH‐Px, the concentration of IgM and the level of AIV‐Ab in the serum.     

Bacteriological evaluation of composted manure solids prepared from anaerobic digested slurry for hygienic recycled bedding materials for dairy cows

Changes in mastitis‐causing pathogens, pH and water content in composted manure solids (CMS) prepared from digested slurry were evaluated during turning at 2‐day intervals for 8 days (C1–C4). The numbers of streptococci, coagulase‐negative staphylococci and coliforms were 2.6 × 10¹, 1.7 × 10² and 1.0 × 10¹ colony‐forming units (cfu)/g in CMS (C4) (summer), and these counts were markedly lower (P < 0.05) than those in CMS (C0 and C1). The bacterial counts ranged from 10¹ to 1.7 × 10² cfu/g in CMS (C4) (summer) and were within approved levels, <1 × 10⁶ cfu/g, indicating a minimal mastitis risk. The temperatures in CMS (C1–C4) increased to 63°C–74°C in summer and 67°C–70°C in winter. The mean pH values in CMS (C0–C4) were 9.2 in summer and 8.7 in winter, and water contents ranged from 61.7% to 69.6% in summer and 73.2% to 66.2% in winter. The significant decrease of pathogenic bacteria in CMS appears to be closely related to temperature >63°C for 8 days, pH 8.7–9.2, and water content 62% to 73%. This study demonstrates that prepared CMS has value as a recycled material with the potential to alleviate udder health issues in dairy cows.     

Feed and water deprivation has a negative but transient effect on the rumen kinetics of Bos indicus steers

The effects on rumen kinetics after feed and water had been deprived for 72 hr were studied using four fistulated Bos indicus steers. The animals were assigned in a 2 × 4 crossover design with two treatments: feed and water ad libitum (control) and no feed and water for 72 hr (deprived) with four steers per treatment over two time periods. Feed and water deprivation caused decreases in the numbers of cellulolytic bacteria (1.4 vs. 0.4 cfu × 10⁶/ml; p = .001), live (23.7 vs. 0.8 × 10⁹/ml; p = .001), dead (12.7 vs. 0.5 × 10⁹/ml; p = .001) and total bacterial counts (36.4 vs. 1.4 × 10⁹/ml; p = .001) at day 0, compared with the control treatment. However, the deprived group had greater numbers of cellulolytic bacteria (2.7 vs. 50.1 cfu × 10⁶/ml; p = .001), live (18.3 vs. 42.2 × 10⁹/ml; p = .001), dead (6. 5 vs. 19.1 × 10⁹/ml; p = .001) and total bacterial counts (24.8 vs. 61.3 × 10⁹/ml; p = .001) from rumen fluid on day 4, compared with the control treatment. The numbers of protozoa in rumen fluid from the deprived group were less than (551.2 vs. 2.4 × 10³/ml; p = .001) the control group on day 0. However, the deprived treatment had fewer protozoa in rumen fluid than the control treatment on day 4 (p = .001) and day 9 (p = .001). Volatile fatty acids and in vitro gas production as functional measurements of rumen fluid followed the same trend as the bacterial and protozoa populations. These results indicate that feed and water deprivation would have a negative but transient effect on the rumen kinetics of Bos indicus steers.     

Potential effect of the microbial fermented feed utilization on physicochemical traits,antioxidant enzyme and trace mineral analysis in rabbit meat

The study investigated the potential effect of the microbial fermented feed utilization on physicochemical traits, antioxidant enzyme and trace mineral analysis in rabbit meat. A total of 72 six-week-old male rabbits were weighed and randomly divided into four groups (1) (SRKC) control; (2) (SRKP) Lactobacillus plantarum 1 × 10⁶ cfu/g fresh weight (FW); (3) (SRKG) Pediococcus acidilactici 1 × 10⁶cfu/g FW and (4) (SRKPG) P. acidilactici + L. plantarum 1 × 10⁶ cfu/g FW. Performance characteristic, weekly body weight, was positively (p < .05) enhanced, while daily feed intake (DFI) and feed convention ratio (FCR) were not influenced in treatments group as compared to untreated. The water, protein, water holding capacity (WHC) and dry matter (DM) concentration were positively (p < .05) influenced, while ash, pH, lightness, redness and yellowness were not influenced in treated group as compared to untreated. The concentration of glutathione peroxidase (Gpx), superoxide dismutase (SOD) and aspartate aminotransferase (AST) was positively (p < .05) influenced in treatments group as compared to control. Regarding trace minerals, copper (Cu), iron (Fe), manganese (Mn) and zinc (Zn) were positively (p > .05) reduced in treated group as compared to untreated. It is concluded that the addition of lactic acid bacteria (L. plantarum and P. acidilactici) in Hybrid pennisetum silage had a constructive influence on rabbit health performance and meat biochemistry.     

Effect of probiotic supplementation on growth performance,intestinal morphology,barrier integrity,and inflammatory response in broilers subjected to cyclic heat stress

This study investigated the protective effects of probiotic on heat stress‐induced intestinal injury and inflammatory response in broilers. A total of 180 male broilers were randomly allocated to three treatments with four replicates each from 22 to 42 days of age. The broilers were either raised under thermoneutral (TN) conditions (23 ± 1°C) or subjected to cyclic heat stress (28–35–28°C for 12 hr daily). The broilers kept at TN conditions were fed a basal diet, and those exposed to heat stress were fed basal diets supplemented with or without probiotic at a dose of 1.5 × 10⁸ cfu/kg. Compared with the TN group, heat stress decreased (p < .05) the growth performance, reduced (p < .05) villus height and villus height: crypt depth ratio in intestinal mucosa, increased (p < .05) serum levels of D‐lactic acid on day 28 and endotoxin, TNF‐α and IL‐6 on day 42, and decreased (p < .05) serum IL‐10 content on day 42. Dietary supplementation of probiotic reversed (p < .05) all these changes except for the growth performance in heat‐stressed broilers. In conclusion, dietary inclusion of probiotic could improve intestinal morphology and barrier function, alleviate inflammatory response, but exert no ameliorative effect on growth performance of broilers under cyclic heat stress.     

Comparison of single- and multi-strain probiotics effects on broiler breeder performance,egg production,egg quality and hatchability

ABSTRACT

1. This study was conducted to investigate the effect of multi-strain probiotic (containing Lactobacillus acidophilus 2.5 × 10⁷ cfu/g, Lactobacillus casei 2.5 × 10⁷ cfu/g, Bifidobacterium thermophilum 2.5 × 10⁷ cfu/g and Enterococcus faecium 2.5 × 10⁷ cfu/g) and single-strain probiotic (Pediococcus acidilactici 1 × 10¹⁰ cfu/g) on broiler breeder performance and gastrointestinal health.

2. A completely randomised trial was conducted using 300 broiler breeder hens (Ross 308) aged 51 weeks old which were randomly allocated to 1 of 5 dietary treatments with 6 replicates per treatment in a 10 week trial. Treatments included (1) the basal diet a negative control, (2) basal diet supplemented with 0.1 g/kg multi-strain probiotic (MS), (3) basal diet supplemented with 0.1 g/kg single-strain probiotic (SS), (4) basal diet supplemented with 0.1 g/kg of both of probiotics (MS+ SS) and (5) positive control basal diet supplemented with 0.5 g/kg oxytetracycline antibiotic (OX).

3. Body weight, egg production, yolk weight, eggshell thickness and weight, Haugh unit, fertility and hatchability were determined. Results showed that dietary treatments had no significant effect on total hen house or total hatching egg production, egg weight, yolk colour index, shell weight, mortality, body weight, fertility, hatchability, oviduct and stroma weight or number of large and small yellow follicles (P > 0.05). None of the jejunum morphological parameters, apparent ileal digestibility of protein and ileal Lactobacillus population were influenced by supplemental probiotics (P > 0.05), although ileum Escherichia coli count was reduced by inclusion of dietary probiotics (P < 0.05).

4. It was concluded that although both probiotic treatments reduced coliforms, they did not improve broiler breeder performance or gastrointestinal tract (GIT) function.     

Effects of butyrate supplementation in antibiotic‐free milk replacer and starter on growth performance in suckling calves

The aim of this study was to evaluate butyrate supplementation of antibiotic‐free milk replacer and starter on growth performance in male Holstein calves. Twenty‐nine calves were divided into two groups. Group C (n = 13) was fed antibiotic‐free milk replacer without supplementation, and Group B (n = 16) was fed antibiotic‐free milk replacer supplemented with butyrate (1.6 % DM of Gustor BP70^®). Starter in Group B contained 0.3 % DM of Gustor BP70^®. The intake of milk replacer was lower in group B than in C (p = 0.07 for the treatment x week interaction). Body weight (BW) and heart girth (HG) in group B was higher than in C during the experimental period (p = 0.07 and 0.01 for the treatment × week interaction, respectively). The duration of the weaning period in group B was shorter than in group C (p = 0.02). β‐hydroxybutyrate (BHBA) was higher in group B than in C (p = 0.04). Insulin like growth factor‐1 (IGF‐1) concentrations tended to be higher in group B than in C (p = 0.07 for treatment × week interaction). Our results show that butyrate supplementation in antibiotic‐free milk replacer and starter exerted positive effects on growth performance in suckling calves.     

Effect of Different Levels of Glycerol and Cholesterol‐Loaded Cyclodextrin on Cryosurvival of Ram Spermatozoa

This study was conducted to determine the optimum level of glycerol and cholesterol‐loaded cyclodextrin (CLC) in a Tris‐based diluent for cryopreservation of ram spermatozoa. Ram semen was treated with 0, 1.5, 3 or 4.5 mg CLC/120 × 10⁶ cells in Tris‐based diluents containing 3, 5 or 7% glycerol in a factorial arrangement 3 × 4 and frozen in liquid nitrogen vapour. Sperm motility, viability (eosin–nigrosin staining) and functional membrane integrity (hypo‐osmotic swelling test) were assessed immediately after thawing (0 h) and subsequently after 3 and 6 h at 37°C. There was an interaction between CLC and glycerol on the functional membrane integrity (p < 0.05). In the presence of 3% glycerol, the highest functional membrane integrity (32.2%) was found in the spermatozoa treated with 1.5 mg CLC/120 × 10⁶ sperm. Post‐thaw sperm motility was highest in 1.5 mg CLC immediately after thawing (40.5%) and after 3‐h (30.6%) incubation at 37°C (p < 0.05). Viability of spermatozoa was higher in all CLC treatments than in the untreated samples, and it was highest (33.9%) in the spermatozoa treated with 1.5 mg CLC (p < 0.05). These data indicate that the addition of cholesterol to sperm membranes by 1.5 mg CLC/120 × 10⁶ cells may allow the use of a lower concentration of glycerol (3%), which is sufficient to mitigate the detrimental effects of freezing and thawing.     

Comparison between the effects of feeding copper sulphate-treated and untreated rapeseed cake containing high glucosinolates on rumen fermentation,nutrient digestion and nitrogen metabolism in steers

Two trials were carried out to study the effects of copper sulphate (CuSO₄) on detoxifying glucosinolates (GLS) in rapeseed cake (RSC) and compare the effects of feeding CuSO₄-treated and untreated RSC on nutrient digestion and nitrogen (N) metabolism in steers. In Trial 1, different concentrations of CuSO₄ solution (1.6 vs. 3.2 g CuSO₄·5H₂O L⁻¹), soaking temperatures (25 vs. 60°C) and drying methods (air drying at 60°C vs. freeze drying) were allocated in a 2 × 2 × 2 factorial arrangement in vitro. In Trial 2, six steers and dietary inclusions of untreated RSC (control), CuSO₄-treated RSC and CuSO₄-added RSC were assigned in a replicated 3 × 3 Latin square design. CuSO₄ treatment in vitro decreased the contents of GLS and thiocyanate (TC) in RSC (p < 0.001). The total amount of GLS and TC decreased by 62.7–68.5% for all treatments. The animal trial showed that CuSO₄-treated RSC inclusion decreased ruminal concentration of valerate (p < 0.01), whereas it did not affect ruminal pH, ammonia N and total volatile fatty acids. Compared with the control, feeding CuSO₄-treated or CuSO₄-added RSC had no effect on plasma concentrations of triiodothyronine and thyroxine, N excretion and N retention. CuSO₄-treated RSC tended to increase neutral detergent fibre digestibility (p = 0.072) and urinary excretion of urea (p = 0.056). Urinary excretion of purine derivatives (p = 0.076) and rumen microbial N supply (p = 0.084) tended to decrease when feeding CuSO₄-treated RSC versus control. TC was found to be the only metabolite of GLS in rumen fluid, plasma and urine. It was feasible to detoxify GLS in RSC using low CuSO₄ at room temperature. However, feeding CuSO₄-treated or CuSO₄-added RSC had minor effects on rumen fermentation, nutrient digestion and N metabolism in steers. CuSO₄ treatment on RSC for feeding steers seems to be unnecessary.     

Effect of laboratory‐isolated Lactobacillus plantarum LGFCP4 from gastrointestinal tract of guinea fowl on growth performance,carcass traits,intestinal histomorphometry and gastrointestinal microflora population in broiler chicken

The study aimed to investigate the effect of feed supplements, viz Lactobacillus plantarum LGFCP4 (laboratory isolate from GIT of Guinea fowl), Lactobacillus acidophilus (NCDC, Karnal) and in‐feed antibiotic bacitracin methylene disalicylate (BMD) on growth performance, FCR, carcass traits and immune organs weight, intestinal histomorphometry and gastrointestinal microflora population in broiler chickens. In a completely randomized design, CARIBRO‐Dhanraja broiler chicks (n = 160) were used with four treatment groups. During the entire experimental duration of 35 days, treatment groups were provided with different dietary treatments (T1 – basal diet (negative control), T2 – antibiotic growth promoter BMD 20 g/100 kg feed (positive control), T3 – 1 × 10⁸ cfu of L. acidophilus/gm‐fermented feed +MOS 1 g/kg feed and T4 – 1 × 10⁸ cfu of laboratory‐isolated L. plantarum LGFCP4/gm‐fermented feed+ MOS 1 g/kg feed. After 35 days of experimental period, no significant results have been observed in different growth performance traits among treatment groups. Cut‐up parts and edible organs' weight remained unaffected by dietary supplementation, whereas weight of immune organs were significantly higher (p < 0.05) in L. plantarum LGFCP4‐supplemented group. At the end of feeding trial, significantly (p < 0.05) lower E. coli count was observed in crop of T4 birds, while in ileum, T2 and T3 showed lower count. In caeca, T2 group showed lowest E. coli count. Salmonella count in crop and ileum was significantly (p < 0.05) low in T3 and T4, while in caeca, T2 group showed lowest count. In terms of histomorphometry, duodenal villous height (VH), crypt depth (CD) and VH:CD ratio were higher for T3 and T4 and lowest values were obtained for T2 group. The results of the study showed that L. plantarum LGFCP4 isolated from GIT of guinea fowl can effectively replace in‐feed antibiotic growth promoters in broiler diets by altering intestinal villi morphology and improving the gut health by reducing the pathogenic microbial load.     

Quantitative and Qualitative Bacterial Flora of Giant Freshwater Prawn Macrobrachium rosenbergii Cultured in Earthen Ponds in Saudi Arabia

Abstract

Quantitative and qualitative analyses of bacterial flora associated with the digestive tract of the giant freshwater prawn Macrobrachium rosenbergii cultured in earthen ponds of Saudi Arabia were carried out. Bacterial counts and flora of prawn-culture pond water, sediment, and prawn carapaces, along with important physicochemical parameters, were investigated, and the isolates were identified to the genus or species level. Total viable counts (TVC; mean ± SD) varied between (1.0 ± 2.3) × 10⁴ and (1.7 ± 0.8) × 10⁵ colony-forming units (cfu) per milliliter in pond water, (5.8 ± 1.7) × 10⁷ and (1.1 ± 2.9) × 10⁹ cfu/g in sediment, (1.5 ± 0.9) × 10⁵ and (1.6 ± 2.4) × 10⁶ cfu/cm² in the carapace, and (9.1 ± 1.5) × 10⁶ and (8.7 ± 1.8) × 10⁷ cfu/g in the digestive tract of freshwater prawns. The bacterial flora was predominantly gram-negative, accounting for 80% of total isolated strains. Altogether, 21 bacterial species of 16 genera were identified. Aeromonas hydrophila, Shewanella putrefaciens, other Aeromonas spp., Vibrio spp., and Enterococcus spp. were the most abundant bacterial species (prevalence ≥10%) in pond water; A. hydrophila, S. putrefaciens, Vibrio spp., and Pseudomonas spp. were the most abundant in sediment; S. putrefaciens, A. hydrophila, Vibrio spp., Enterococcus spp., and Aeromonas spp. were the most abundant on the prawn carapace; and A. hydrophila, S. putrefaciens, Vibrio spp., Enterococcus spp., and Pasteurella spp. were the most abundant in the digestive tracts. In every population studied, Aeromonas spp., S. putrefaciens, Enterococcus spp., Vibrio spp., Pasteurella spp., Chryseomonas spp., and Pseudomonas spp. were present.     

Pharmacokinetic–pharmacodynamic relationship of marbofloxacin for Escherichia coli and Pasturella multocida following repeated intramuscular administration in goats

The pharmacokinetics (PK) and pharmacodynamics (PD) of marbofloxacin (MBF) were determined in six healthy female goats of age 1.00–1.25 years after repeated administration of MBF. The MBF was administered intramuscularly (IM) at 2 mg kg^?1 day^?1 for 5 days. Plasma concentrations of MBF were determined by high‐performance liquid chromatography, and PK parameters were obtained using noncompartmental analysis. The MBF concentrations peaked at 1 hr, and peak concentration (C_max) was 1.760 µg/ml on day 1 and 1.817 µg/ml on day 5. Repeated dosing of MBF caused no significant change in PK parameters except area under curve (AUC) between day 1 (AUC_0–∞D1 = 7.67 ± 0.719 µg × hr/ml) and day 5 (AUC_0‐∞D5 = 8.70 ± 0.857 µg × hr/ml). A slight difference in mean residence time between 1st and 5th day of administration and accumulation index (AI = 1.13 ± 0.017) suggested lack of drug accumulation following repeated IM administration up to 5 days. Minimum inhibitory concentration (MIC) demonstrated that Escherichia coli (MIC = 0.04 µg/ml) and Pasturella multocida (MIC = 0.05 µg/ml) were highly sensitive to MBF. Time‐kill kinetics demonstrated rapid and concentration‐dependent activity of MBF against these pathogens. PK/PD integration of data for E. coli and P. multocida, using efficacy indices: C_max/MIC and AUC_0–24hr/MIC, suggested that IM administration of MBF at a dose of 2 mg kg^?1 day^?1 is appropriate to treat infections caused by E. coli. However, a dose of 5 mg kg^?1 day^?1 is recommended to treat pneumonia caused by P. multocida in goats. The study indicated that MBF can be used repeatedly at dosage of 2 mg/kg in goats without risk of drug accumulation up to 5 days.     

Pharmacokinetics and pharmacodynamics of alfaxalone after a single intramuscular or intravascular injection in mallard ducks (Anas platyrhynchos)

Pharmacokinetics and pharmacodynamics of alfaxalone was performed in mallard ducks (Anas platyrhynchos) after single bolus injections of 10 mg/kg administered intramuscularly (IM; n = 10) or intravenously (IV; n = 10), in a randomized cross‐over design with a washout period between doses. Mean (±SD) C_max following IM injection was 1.6 (±0.8) µg/ml with T_max at 15.0 (±10.5) min. Area under the curve (AUC) was 84.66 and 104.58 min*mg/ml following IV and IM administration, respectively. Volume of distribution (V_D) after IV dose was 3.0 L/kg. The mean plasma clearance after 10 mg/kg IV was 139.5 (±67.9) ml min^?1 kg^?1. Elimination half‐lives (mean [±SD]) were 15.0 and 16.1 (±3.0) min following IV and IM administration, respectively. Mean bioavailability at 10 mg/kg IM was 108.6%. None of the ducks achieved a sufficient anesthetic depth for invasive procedures, such as surgery, to be performed. Heart and respiratory rates measured after administration remained stable, but many ducks were hyperexcitable during recovery. Based on sedation levels and duration, alfaxalone administered at dosages of 10 mg/kg IV or IM in mallard ducks does not induce clinically acceptable anesthesia.     

A low protein maternal diet during gestation has negative effects on male fertility markers in rats – A Systematic Review and Meta-analysis

Research indicates that some adult diseases including reproductive pathologies are programmed in utero during foetal development. In particular, maternal low dietary protein, during the most critical developmental periods of male foetal development, may have a detrimental impact on male fertility through direct and epigenetic mechanisms. The aim of our study was to evaluate the impact of a gestational low protein diet on fertility markers in male offspring in rats through a systematic review and meta-analysis. A systematic search using PubMed, and EMBASE databases was performed and two investigators independently screened the 1,703 prospective articles. Eleven articles met the eligibility criteria. Outcome measures were pooled using random-effects models and expressed as mean differences (MDs) at 95% CIs for each study. The results reveal significant reduction in testis weight (MD (mean difference) −0.08 g; −0.12, −0.42; p = .0001), epididymal sperm count (MD −35.34 × 10⁶ cells; −52.15, −18.53; p = .0001), number of Sertoli cells (MD −7.27 × 10⁶ (−13.92, −0.62; p = .03), testosterone (T) concentration (MD −0.29 ng/ml; −0.48, −0.09; p = .004) and luteinising hormone (LH) concentration (MD of −0.24 ng/ml; −0.45, 0.04; p = .02) in comparison with controls. In contrast, follicle-stimulating hormone (FSH) concentration (MD of 0.07 ng/ml; −0.16, 0.29; p = .56) was not significantly different from controls. We conclude that low gestational dietary protein maternal intake potentially negatively impacts fertility in male progeny later in life. The mechanisms of action responsible for these phenomena remain unclear.     

Factors influencing production of hygienic raw milk by small scale dairy producers in selected areas of the Jaffna district,Sri Lanka

The objective of the study was to investigate the influence of dairy cow management techniques and milking methods on hygienic quality of raw milk. Total Bacterial Count (TBC) and Total Coliform Colonies (TCC) were studied to determine the effects. Investigations were carried out in fifty dairy farms from August 2007 to December 2007. The mean TBC and TCC for the herds with comparatively good and poor management practices were 0.9 × 10⁵ cfu/ml and 0.2 × 10³/ml and 99 × 10⁵ cfu/ml and >180 × 10³/ml, respectively. The overall mean TBC (22 × 10⁵ cfu/ml) and TCC (47 × 10³/ml) obtained in this study exceeded the internationally recommended levels for TBC (10⁵ cfu/ml) and TCC (<1,000/ml). The overall results obtained suggested that the raw milk tested was of poor hygienic quality with the presence of a great variability among milk samples.     

Evaluation of a portable device for assessment of motility in stallion semen

In horse breeding, quality assessment of semen before insemination is often requested. Non‐laboratory‐based techniques for objective analysis of sperm motility are thus of interest. The aim of this study was evaluating a portable device for semen analysis (Ongo sperm test) and its comparison with computer‐assisted semen analysis (CASA). Semen was collected from 10 stallions, diluted to 100, 50 and 25 × 10⁶ sperm/ml and analysed for total (TM) and progressive motility (PM). The final sperm concentration influenced total motility analysed by Ongo (p < 0.05) which was higher at 100 × 10⁶ sperm/ml when compared to 25 × 10⁶ sperm/ml (p < 0.05) but not when compared to 50 × 10⁶ sperm/ml (n.s.). Sperm concentration did not influence total motility when assessed by SpermVision (n.s.). Agreement between methods was evaluated by correlation analysis and Bland–Altman plot. Intra‐assay variation of Ongo was 5.2% ± 3.0 for TM and 6.9% ± 3.4 for PM. Correlation between Ongo and CASA was r = 0.79, 0.88 and 0.83 for 100, 50 and 25 × 10⁶ sperm/ml for TM, and r = 0.87, 0.89 and 0.87 for PM, respectively (all p < 0.001). At the 100 and 25 mio/ml dilutions, the difference between the two systems deviated significantly from 0, while no such bias existed at the 50 mio/ml dilution (TM Ongo 85.0%, CASA 82.3%; PM Ongo 64.1%, CASA 66.1%). The 95% confidence interval was 19.9%, 18.9% and 19.2% ± mean for TM and 20.7%, 17.4% and 20.3% ± mean for 100, 50 and 25 × 10⁶ sperm/ml, respectively. In conclusion, Ongo sperm test sperm motility data were strongly correlated with data obtained by CASA. In addition, at a concentration of 50 × 10⁶ sperm/ml values measured with both systems were close to identical. At this concentration, which is recommended in equine AI, Ongo and CASA can be used interchangeably.     

Total antioxidant capacity and protein peroxidation intensity in seminal plasma of infertile and fertile dogs

The aim of this study was to evaluate the total antioxidant capacity and protein peroxidation intensity in seminal plasma of infertile and fertile dogs. The study was conducted on 10 infertile and 10 fertile dogs of various breeds. Infertility was defined as conception failure at least three matings with different bitches. Semen was collected by manual manipulation. The sperm concentration and motility parameters were evaluated using CASA Hamilton Thorne, Vers. IVOS 12.3. The morphology of spermatozoa and the percentage of live and dead sperm cells were assessed microscopically, total antioxidant capacity and the content of SH‐groups in seminal plasma were determined spectrophotometrically, the contents of protein peroxidation markers in seminal plasma, bityrosine and formylokinurenine, were determined using spectrofluorimetric methods. Sperm concentration and total sperm count were significantly (p < 0.05) lower in infertile dogs than in fertile dogs (99.92 ± 3 0.05 × 10⁶/ml vs. 282.07 ± 48.27 × 10⁶/ml; 214.19 ± 114.74 × 10⁶ vs. 747.57 ± 210.94 × 10⁶, respectively). The percentage of spermatozoa with normal morphology and the most determined motility parameters differed significantly (p < 0.05) between both groups. The mean values of total antioxidant capacity in the seminal plasma were significantly (p < 0.05) lower (19.95 ± 20.94 vs. 25.66 ± 23.18 µmol/g protein), whereas the mean contents of bityrosine and formylokinurenine in seminal plasma were significantly (p < 0.05) higher in infertile dogs than in fertile dogs (3.71 ± 4.83 µg/mg protein vs. 1.55 ± 2.00 µg/mg protein and 0.37 ± 0.45 µg/mg protein vs. 0.14 ± 0.08 µg/mg protein, respectively). In conclusion, the obtained results suggest that the poor semen quality and infertility in dogs could be associated with lowered total antioxidant capacity and increased protein peroxidation in seminal plasma as a consequence of oxidative stress.     

Impact of intramammary tilmicosin infusion as a dry cow therapy

Three hundred subclinically infected quarters of 259 Holstein cows infected with gram‐positive bacteria were selected via quota sampling based on the California Mastitis Test (CMT) result and were divided randomly and equally into treatment and test groups. Quarters of test group (n = 150 in 128 cows) were treated with an intramammary infusion of tilmicosin, and quarters of the control group (n = 150 in 131 cows) were treated with cloxacillin as a traditional intramammary infusion of dry cow (DC) ointment. Cows with more than one infected quarter were randomly assigned to the same group, and adjacent quarters were treated the same. The milk samples of all quarters were obtained, and bacterial cultures and somatic cell count (SCC) were tested before dry cow therapy (DCT) (50 ± 15 days before parturition), and finally on day 2 of the next lactation. Results have shown that total bacteriological cure rates on day 2 of the next lactation were 45% and 78%, (p = .01), new infection rates were 43.3% and 56.6%, and SCC was (6.732 × 10⁵ ± 3.124 × 10⁵) and (5.025 × 10⁵ ± 2.935 × 10⁵), (p > .05) in test and control groups, respectively. Tilmicosin had less effect on reducing IMI due to Corynebacterium bovis, and had no effect on Streptococcus agalactiae, but had a potent effect against Staphylococcus aureus. It was concluded that tilmicosin alone should not be infused as an alternative to conventional dry cow therapy. However, it had a significant effect against S. aureus, and the potential of tilmicosin to treat S. aureus IMI should be confirmed in further studies.     

Effects of chestnut tannins on intestinal morphology,barrier function,pro‐inflammatory cytokine expression,microflora and antioxidant capacity in heat‐stressed broilers

A study was conducted to evaluate the effects of chestnut tannins (CT) on intestinal morphology, barrier function, pro‐inflammatory cytokine expression, microflora and antioxidant capacity in heat‐stressed broilers. Four hundred 28‐day‐old male Ross 308 broilers were randomly assigned into four groups, with 10 replicates per group and 10 broilers per replicate. The broilers in the normal (NOR) group were kept at 22 ± 1°C and fed the basal diet, and each of the other three groups were treated with cyclic heat (33 ± 1°C from 0800 to 1800 and 22 ± 1°C from 1800 to 0800) and fed the basal diet with 0 (HT), 1 (CT1) or 2 (CT2) g of CT/kg of diet. The experiment lasted for 14 days. Compared with the HT group, broilers in the NOR and CT2 groups had higher (p < .05) average daily gain and villus height in the jejunum and lower serum d ‐lactate (p < .001) and diamine oxidase (p < .01) levels. The addition of 2 g CT/kg of diet increased the total antioxidant capacity (p < .001) and superoxide dismutase activities (p < .05) and zonula occludens‐1 mRNA expression level (p < .05) and decreased the malondialdehyde concentration (p < .01) and mRNA expression levels of interleukin‐6 (p < .001) and nuclear factor kappa B (p < .001) in the jejunal mucosa of heat‐stressed broilers. The populations of Escherichia coli and Clostridium in the jejunum (p < .01) and caecum (p < .05) of broilers in the HT group were higher than those in the NOR and CT2 groups. In conclusion, the addition of 2 g CT/kg of diet seemed to be a feasible means of alleviating the negative effects of heat stress on the growth performance and intestinal function of broilers.     

The effect of cooling to different subzero temperatures on dog sperm cryosurvival

The objective was to assess the effect of cooling to different subzero temperatures around ice formation (?5°C) on dog sperm cryosurvival and plasma membrane fluidity. Semen was centrifuged, and sperm were resuspended in a Tris‐egg yolk medium (3% glycerol). Diluted sperm were cooled from 22 to 5°C, and then, a Tris‐egg yolk medium containing 7% glycerol was added (final concentration of 5% glycerol and 200 × 10⁶ cells/ml). Sperm were packaged in 0.5‐ml plastic straws, and equilibration was done 16 hr at 5°C before freezing. I. Straws (n = 47) at 5°C were exposed to nitrogen vapours to determine the freezing point. II. Other straws (from different ejaculates) processed as mentioned, were further cooled to ?3, ?5 or ?7°C and immediately rewarmed in a water bath at 37°C. Motility, plasma membrane functionality and acrosome integrity were assessed. III. Other straws (from different ejaculates) processed as mentioned were further cooled to ?3 or ?5°C, frozen over nitrogen vapours and stored in liquid nitrogen for one month. Straws were thawed in a water bath at 38°C for 30 s. Motility, plasma membrane functionality, plasma membrane integrity, acrosome integrity, capacitation status and plasma membrane fluidity were assessed. Ice nucleation temperature was ?14.3 ± 2.05°C (mean ± SD); cooling to +5, ?3, ?5 and ?7°C, without freezing, produces no differences on sperm quality between target temperatures; cooling to +5, ?3, and ?5°C produced no differences on sperm survival and plasma membrane fluidity after freeze–thawing. In conclusion, cooling of dog spermatozoa to different subzero temperatures did not improve sperm cryosurvival and had no effect on plasma membrane fluidity after thawing.