Understanding the effects of intramuscular injection and feed withdrawal on Salmonella Typhimurium shedding and gut microbiota in pullets

Background:Gut microbiota plays a key role in health,immunity,digestion,and production in layers.Factors such as environment,diet,diseases,stress,and flock management significantly affect gut microbiota;however,it is not known how potential stressors such as intramuscular injections or feed withdrawal alter the composition of gut microbiota that result in increased the shedding level of foodborne pathogens.In the current study,the effects of intramuscular corticosterone injection and feed withdrawal were evaluated to understand their role in Salmonella Typhimurium shedding and changes in the composition of gut microbiota in layers.Results:Salmonella shedding was observed for 8 weeks post-infection.There was a significant increase in Salmonella Typhimurium count after intramuscular injection and feed withdrawal.The Salmonella infected and the negative control groups showed significant differences in the abundance of different genera in gut microbiota at week 1 and up to week 7 post infection.The infected group showed a significant reduction in alpha diversity of gut microbiota.Firmicutes reduced significantly(P<0.05)after intramuscular injection,while the feed withdrawal groups did not cause any significant changes in Proteobacteria-Firmicutes ratio.Furthermore,intramuscular injection resulted in a significant change in alpha diversity of gut microbiota.Conclusions:Exposure of chicks to relatively low dose of Salmonella Typhimurium can lead to persistent shedding in pullets.The Salmonella Typhimurium infection disrupted the gut microbiota composition immediately after infection.The potential stress of intramuscular injection and feed withdrawal significantly increased the Salmonella Typhimurium count in faeces.The intramuscular injection also resulted in a significant alteration of the Proteobacteria-Firmicutes ratio,which could increase the risk of dysbiosis.     

Effect of eCG dose on ovarian haemodynamics,hormonal profiles and prolificacy rate when oestrus was induced during low-breeding season in Beetal goats

The objectives of the experiment were to determine the effect of two doses of equine chorionic gonadotropin (eCG) in a standard synchronization protocol based on a short-term progesterone (P₄) priming on ovarian structures and haemodynamics, concentrations of steroid hormones and prolificacy rate when oestrus was induced during low-breeding season (LBS) in Beetal dairy goats. We hypothesized that inclusion of eCG in a short-term P₄ priming-based synchronization protocol would increase the blood perfusion to ovarian structures leading to enhance oestrous and ovulatory responses and prolificacy rate in goats. Forty-two multiparous acyclic goats were blocked by body condition and, within block, assigned randomly to receive saline as control (CON), low eCG (L-eCG; 300 IU) or high eCG (H-eCG; 600 IU) dose. Initially, a controlled internal drug release (CIDR) device was placed in the anterior vagina on d −8, followed by removal of CIDR on d −3, concurrent with the administration of PGF_2α and eCG according to their respective treatments. Goats were monitored for oestrous response. B-mode and Doppler ultrasonography was performed with 12-h interval, starting from day −3 until natural breeding (day 0), and then on days 5, 10, 15 and 20 post-breeding to monitor follicular and luteal dynamics and blood flow, respectively. Blood was sampled at 0, 12, 24, 36 and 60 h after CIDR removal to quantify plasma concentrations of estradiol-17β (E₂), whereas plasma concentrations of P₄ were assayed at days 5, 10, 15 and 20 after breeding. Pregnancy and prolificacy rates were determined at day 30 and 150 after breeding, respectively. Data were analysed with mixed-effects models, and orthogonal contrasts were used to evaluate the effect of treatment [Con vs. (½ L-eCG + ½ H-eCG)] and dose of eCG (L-eCG vs. H-eCG). Data are presented in sequence as CON, L-eCG, H-eCG (LSM ± SEM). The oestrous intensity score (152.9 vs. 182.7 vs. 186.5 ± 15.1; p = .02) was greater in eCG-treated goats as compared to CON. Administration of eCG reduced the intervals to standing oestrus (66.2 vs. 41.8 vs. 48.9 h ± 5.5; p = .05), breeding (70.2 vs. 44.4 vs. 45.4 h ± 4.5; p = .03) and ovulation (84.5 vs. 61.2 vs. 63.4 h ± 6.2; p = .05) compared with CON goats. The mean growth rate of pre-ovulatory follicle was greater (1.11 vs. 1.49 vs. 1.45 mm ± 0.08; p = .01) in eCG-treated goats resulting in an increased diameter of pre-ovulatory follicle (6.27 vs. 7.20 vs. 7.31 mm ± 0.07; p < .01) and corpora lutea (6.75 vs. 8.26 vs. 8.07 mm ± 0.42; p = .04) than CON. The mean follicular blood flow did not differ among treatments; however, the mean luteal blood flow was greater in L-eCG-treated goats (0.81 vs. 1.61 vs. 1.07 cm² ± 0.12; p = .001). The mean concentrations of E₂ (4.03 vs. 5.21 vs. 4.78 pg/ml ± 0.42; p = .04) and P₄ (4.85 vs. 6.39 vs. 6.22 ng/ml ± 0.34; p = .04) were greater in eCG-treated goats. The twinning rate did not differ between treatments; nevertheless, prolificacy rate was greater (p = .04) in L-eCG-treated goats. Collectively, our data suggest that the administration of eCG improves the induction of oestrous and ovarian dynamics. Administration of L-eCG enhances prolificacy rate, therefore, a low dose of eCG might be practically beneficial to improve reproduction during LBS in acyclic Beetal dairy goats.     

Performance and carcass measurements of ewe lambs reared in a feedlot and fed wheat (<Emphasis Type="BoldItalic">Triticum durum</Emphasis> Desf.) middlings total mixed rations in the summer season

The effect of total mixed ratios containing wheat middlings (WM) as a corn grain substitute on the growth performance and carcass traits of Comisana ewe lambs was evaluated. Forty ewe lambs, with average live body weight of 13 ± 0.3 kg (mean ± SEM), were allocated randomly to two isocaloric and isonitrogenous diets for 50 days. Control diet contained 400 g/kg of dry matter (DM) of corn as the main starchy source, whereas experimental diet contained 600 g/kg DM of WM. In vivo nutrient apparent digestibility of the two diets was determined using Comisana rams (mean body weight, 65 ± 2.3 kg) and indicated significant (P < 0.05) differences for neutral detergent fibre, acid detergent fibre, cellulose and hemicellulose fractions. Results from the trial using Comisana ewe lambs showed that growth traits were unaffected by dietary treatments as well as none of the carcass measurements examined (P > 0.05). These findings indicate that WM results as a suitable feed ingredient for growing ewe lambs that can be a satisfactory substitute to conventional grain source.     

Cross-bred Cattle Protected against Hyalomma anatolicum anatolicum by Larval Antigens Purified by Immunoaffinity Chromatography

Antigens from larvae of Hyalomma anatolicum anatolicum were extracted and purified by immunoaffinity chromatography using immunoglobulin ligands from cross-bred animals immunized with soluble larval antigen. Affinity-purified antigen (Aff-TLE) and a total larval extract (TLE) were used to immunize cross-bred (Bos indicus×Bos taurus) cattle. The group immunized with Aff-TLE rejected 71.6% of larvae and 77.3% of nymphs. However, the rejection percentages were lower in the TLE-immunized group. No significant changes in the feeding period, moulting percentages or moulting period of engorged larvae and nymphs were recorded. There was, however, a significant decrease in the number of resultant nymphs p<0.01) and adults (p<0.01) in the ticks fed on the Aff-TLE-immunized group. The Aff-TLE antigen was 93.3% purified. SDS-PAGE analysis identified a 39 kDa protein, reported for the first time, as the antigen responsible for the induction of resistance in the host.     

Evaluation of Plasmid Delivery by Electroporation as a Means of Increasing Gonadotropin-Releasing Hormone Production in Stallions

A plasmid delivery system validated in other species was assessed for its potential for inducing long-term expression of gonadotropin-releasing hormone (GnRH) in stallions. The efficacy of this technique was demonstrated using two plasmids: pSEAP, expressing secreted embryonic alkaline phosphatase (SEAP), and pGnRH, expressing GnRH. In experiment 1, geldings were used as a model to test the effect of muscle of injection (splenius, pectoralis, and semitendinosus; n = 3 for each site) on the expression of the reporter plasmid, pSEAP. Concentrations of SEAP rose (P < .01) in jugular plasma samples, indicating uptake and expression of the pSEAP plasmid. Concentrations of SEAP were greatest (P < .05) and most consistent after pectoralis injection, and this site was chosen for injection and electroporation in the subsequent experiment. In ex-periment 2, stallions were treated with pGnRH (2 mg, n = 3; and 4 mg, n = 3) or 2 mg of pSEAP (control; n = 4) to determine the effects on the reproductive axis. Treatment with pGnRH (day 0) resulted in higher (P < .05) plasma testosterone concentrations from day 35 to 56 and increased the luteinizing hormone (LH) (P < 0.01) and testosterone (P < .1) responses to GnRH challenge on day 21. Daily semen characteristics from days 31 to 36 showed no effect (P > .1) of pGnRH treatment on seminal characteristics. It was concluded that delivery by electroporation of plasmids encoding peptide hormones may serve as a means of long-term in vivo production of peptides in the horse. Increases in LH and testosterone secretion after GnRH were observed in pGnRH-treated stallions; however, optimal conditions for expression need to be determined in future experiments.     

Endometrial nitric oxide synthase activity in mares susceptible or resistant to persistent breeding‐induced endometritis and the effect of a specific iNOS inhibitor in vitro

Emerging research suggests that the nitric oxide system may play a role in persistent breeding‐induced endometritis (PBIE) in the mare. Differences in uterine nitric oxide (NO) levels between mares susceptible or resistant to PBIE and a dose‐dependent inhibitory effect of NO on uterine contractility have been demonstrated. The objectives of this study were to investigate the difference in total nitric oxide synthase (NOS) activity of the endometrium between susceptible and resistant mares and the effect of a specific inducible nitric oxide synthase (iNOS) inhibitor on the endometrial NOS activity in vitro. Six susceptible and six resistant mares were selected based on preset criteria and the results of an intrauterine challenge with killed spermatozoa during oestrus. Endometrial biopsy samples were collected 24 hr post‐challenge and cultured at 37°C for 24 hr in L‐arginine supplemented minimum essential medium with or without a specific iNOS inhibitor (1,400 W dihydrochloride, 1 mM). The medium and the cultured endometrial tissue were collected after 24 hr of culture and assayed for NO and total protein, respectively. Total NO content of the medium, normalized to endometrial tissue wet weight or total protein, was used as a measure of endometrial NOS activity. Non‐parametric tests were applied for statistical analysis. Susceptible mares had significantly greater endometrial NOS activity than resistant mares. The iNOS inhibitor treatment significantly reduced NOS activity in endometrial samples derived from susceptible and resistant mares. These findings provide a basis for in vivo testing of specific iNOS inhibitors as preventative or therapeutic options for PBIE in mares.     

Modification and characterization of biodegradable methylcellulose films with trimethylolpropane trimethacrylate (TMPTMA) by γ radiation: effect of nanocrystalline cellulose

Methylcellulose (MC)-based films were prepared by solution casting from its 1% aqueous suspension containing 0.25% glycerol. Trimethylolpropane trimethacrylate (TMPTMA) monomer (0.1-2% by wt) along with the glycerol was added to the MC suspension. The films were cast and irradiated from a radiation dose varied from 0.1 to 10 kGy. Then the mechanical properties such as tensile strength (TS), tensile modulus (TM), and elongation at break (Eb) and barrier properties of the films were evaluated. The highest TS (47.88 PMa) and TM (1791.50 MPa) of the films were found by using 0.1% monomer at 5 kGy dose. The lowest water vapor permeability (WVP) of the films was found to be 5.57 g·mm/m(2)·day·kPa (at 0.1% monomer and 5 kGy dose), which is 12.14% lower than control MC-based films. Molecular interactions due to incorporation of TMPTMA were supported by FTIR spectroscopy. A band at 1720 cm(-1) was observed due to the addition of TMPTMA in MC-based films, which indicated the typical (C═O) carbonyl stretching. For the further improvement of the mechanical and barrier properties of the film, 0.025-1% nanocrystalline cellulose (NCC) was added to the MC-based suspension containing 1% TMPTMA. Addition of NCC led to a significant improvement in the mechanical and barrier properties. The novelty of this investigation was to graft insoluble monomer using γ radiation with MC-based films and use of biodegradable NCC as the reinforcing agent.     

Possible role of central interleukins on the anorexigenic effect of lipopolysaccharide in chicks

1. The purpose of the present study was to determine if central interleukin-1β (IL1β), interleukin-6 (IL6) and interleukin-8 (IL8) affect feeding behaviour in chicks (Gallus gallus) and examine if central interleukins are related to the lipopolysaccharide (LPS)-induced anorexia.

2. Intra-abdominal (IA) injection of LPS significantly suppressed feeding behaviour and significantly increased mRNA expression of IL1β and IL8 in the diencephalon when compared to the control group, while IL6 tended to be increased.

3. Intracerebroventricular (ICV) injection of 200 ng IL1β significantly decreased food intake at 60 min after the injection while IL6 and IL8 had no effect.

4. IA injection of these ILs (200 ng) had no effect on food intake in chicks.

5. ICV injection of 200 ng IL1β did not affect water intake and plasma corticosterone concentration, suggesting that central IL1β might not be related to the regulation of drinking behaviour and the hypothalamus–pituitary–adrenal axis.

6. The present study demonstrated that central IL1β but not IL6 and IL8 might be related to the inhibition of feeding in chicks.     

An immunochromatographic serological assay for the diagnosis of Mycobacterium tuberculosis

A rapid serological test for tuberculosis (TB) infection was designed using antigens specific to Mycobacterium tuberculosis. Tuberculosis infection, TB vaccination and exposure to environmental Mycobacteria cannot be distinguished using skin tests based on tuberculin protein derivatives. The standard diagnostic techniques such as skin tests, X-rays and DNA techniques are time consuming, expensive, and not practical for screening large populations. We used the 38, 63, 64, 14, 59-kDa antigens of M. tuberculosis to develop a rapid immunochromatographic test kit. This study evaluates the diagnostic potential of the rapid test kit using TB positive and TB negative serum samples from various hospitals in India. The samples were obtained from patients infected with or exposed to bacteria and viral pathogens. The results demonstrated that the combination of antigens improved the diagnostic specificity and sensitivity. The specificity of the test was 99.42% with sensitivity of 98.52% (n = 241). In case of multiple infections, the specificity was 93.15% with a low sensitivity of 73.52% n = 141). The test kit may offer an improved alternative to purified protein derivative (PPD). This rapid TB test kit may be a useful tool for first-line testing of suspected cases, epidemiological studies and in designing a quality health system to reduce health hazards in resource-poor countries.