Use of Penicillin and Streptomycin to Reduce Spread of Bacterial Coldwater Disease I: Antibiotics in Sperm Extenders

Bacterial coldwater disease caused by Flavobacterium psychrophilum has led to the loss of significant numbers of hatchery-reared salmonids. The bacteria can be spread from parent to progeny within contaminated sperm and ovarian fluid. Methods for disinfecting ovarian fluid and unfertilized eggs are available, but methods for disinfecting sperm have not been described. In this study we determined whether sperm extenders containing a mixture of penicillin and streptomycin can be used to eliminate F. psychrophilum. In vitro trials demonstrated that when Rainbow Trout Oncorhynchus mykiss sperm is mixed with an extender, a 15-min exposure to 0.197 mg penicillin plus 0.313 mg/mL streptomycin is effective at killing the bacteria and has no effect on sperm motility. Small-scale trials showed that egg fertilization rates were not reduced when sperm held in an extender solution containing the same antibiotic mixture for 15 min was used to fertilize eggs. Production-scale trials, however, showed a roughly 18% decrease in egg fertilization rate when sperm stored in an antibiotic containing extender was used. To determine why a reduction in fertilization capacity was observed, a small-scale experiment testing the fertilization of eggs with larger quantities of sperm was performed and showed that increasing the volume of sperm used did not increase fertilization rates. Our results demonstrate that extenders containing penicillin and streptomycin can be used to disinfect sperm, especially when small quantities of eggs are fertilized, but factors negatively affecting egg fertilization and survival on a production scale still need further investigation.

Received May 1, 2014; accepted August 10, 2014     

Bacterial Analysis of Fertilized Eggs of Atlantic Salmon from the Penobscot,Naraguagus, and Machias Rivers,Maine

Serious losses have occurred at the U.S. Fish and Wildlife Service, Craig Brook National Fish Hatchery, East Orland, Maine, among eggs that were taken from Atlantic Salmon Salmo salar, which were held as captive broodfish during their returns to the Penobscot River, Naraguagus River, and Machias River to spawn. Bacterial isolations were attempted from external surfaces and the internal contents of individual eggs. Externally and in all cases, Pseudomonas fluorescens was the predominant bacterium associated with the surface of all eggs. These bacteria were resistant to a surface treatment of 1,667 ppm formalin for 15 min and, therefore, the monoclonal nature of P. fluorescens on egg surfaces was considered to result from its ability to resist the germicidal activity of formalin administered for antifungal treatments. Flavobacterium psychrophilum, the cause of bacterial coldwater disease, was isolated from the interior of 23.6, 18.1, and 29.2% of the dead Atlantic Salmon eggs from Penobscot River egg lots A-98, A-100, and A-101, respectively, and concentrations of this pathogen ranged from 1.0 × 10³ to >5 × 10⁸ CFU per gram of dead egg. Flavobacterium psychrophilum was also isolated from 8.3, 26.7, and 10.0% of the dead eggs from Naraguagus River egg lots N-158, N-161, and N-163, respectively, in which concentrations of this organism ranged from 1.0 × 10³ to 7.5 × 10⁸ CFU per gram of egg. This bacterium was also isolated from within 18.3% and 3.3% of the dead eggs from Machias River egg lots M-128 and M-142, respectively, and its concentrations ranged from 1.0 × 10³ to 1.5 × 10⁸ CFU per gram of egg. The finding of F. psychrophilum from within these eggs is indicative of this pathogen's widespread and persistent prevalence in Atlantic Salmon in New England.

Received December 19, 2014; accepted May 4, 2015     

Antimicrobial substances and chick growth promotion: Comparative studies on selected compounds in vitro and in vivo

1. Antibacterial activity of selected compounds in vitro and their ability to promote growth when added to the diet of chicks were not correlated.

2. The difference in growth‐promoting activity between streptomycin, which was active, and kanamycin which was not, was not related to their effects on the flora adhering to the wall of the crop, jejunum or ileum, to their relative toxicity to the chick nor to differences in their stability in the diet.

3. When low concentrations ofkanamycin or streptomycin were given, the antibiotic was concentrated in the caecal contents. In contrast, penicillin was undetectable in the caeca of birds given high concentrations of benzyl penicillin.

4. In chicks given benzyl penicillin, the numbers of Streptococcus faecium increased at the expense of Strep, faecalis. This probably resulted from changes in the crop flora.

5. Birds given high dietary concentrations of benzyl penicillin showed improved weight gains, in spite of increased numbers of Escherichia coli in the small intestine.     

Effect on fertility of storing turkey semen for 24 hours at 10 °C in fluids of different pH

1. A study was undertaken into the storage of turkey semen in vitro for 24 h.

2. Turkey semen storage for 24 h at 10 °C was not noticeably improved by using a buffered diluent compared with an unbuffered (pH 7.3) diluent. This is different from the case of the fowl and probably reflects differences in the metabolism of spermatozoa between the species.

3. Adverse effects on the fertilising ability of turkey spermatozoa were observed with diluents buffered around pH 7.4 and 5.8 when compared with the use of diluent either buffered at pH 7.1 or unbuffered.     

The effect of tonicity of storage media for fowl semen on the occurrence of neck‐bending spermatozoa,fertility and hatchability 1

1. Semen of White Leghorn and Rhode Island Red males was stored for 24 h in diluents hypertonic (460 mOsm/kg H₂O) and isotonic (340 mOsm/kg H₂O) to cock seminal plasma.

2. Compared with the fertility results with semen that had been stored in the hypertonic diluent or was fresh, the fertility of the White Leghorns was not affected after storage in the isotonic diluent; a decrease (P < 0.lb05) was observed, however, using Rhode Island Red semen and isotonic diluent.

3. Fresh RIR semen contained 2.lb31% “neck‐bent spermatozoa” (NBS) which was increased to 4.lb23% and 5.lb76% after dilution in hypertonic and isotonic diluents respectively and stored for 24 h. It is doubtful whether this increase (P < 0.lb05) is the sole reason for the lowered fertility obtained with this breed after storage in the isotonic diluent.     

Hatchability rate and embryonic growth of broiler chicks following in ovo injection royal jelly

1. The objectives of this study were to compare the hatchability, chick body and internal organs weights and plasma testosterone concentration of hatchlings after in ovo administration of royal jelly (RJ) on d 7 of incubation.

2. Fertile eggs (n = 150) were injected into the air sac or yolk sac with 0.5 ml normal saline solution or normal saline and pure RJ. The eggs were randomly divided into 5 groups of 30 eggs each: NC, control eggs receiving no injection; ASA, air sac–injected eggs given normal saline solution; ARJ, air sac–injected eggs injected with pure RJ; YSA, yolk sac–injected eggs receiving normal saline solution and YRJ, yolk sac–injected eggs given pure RJ.

3. Injection of RJ significantly decreased hatchability (46.7%) compared with injection of saline solution (68.3%). Hatchability was lower in ARJ (33.3 %) and YRJ (60.0%) groups than in the NC group (90.0%). Hatchability in ASA (70.0%) and YSA (66.66%) groups were comparable to the NC group.

4. In ovo injection of RJ into both sacs increased chicks’ absolute and relative body, heart, liver and testes weights compared to the control group whereas plasma testosterone concentration was similar among the different groups.

5. It was concluded that in ovo injection of RJ may be an effective method to increase the body weight of hatched chicks as an absolute value (CWT) and chicks' internal organ weights.     

Studies on occurrence,characterisation and decontamination of emerging pathogenic Escherichia coli (STEC,ETEC and EIEC) in table eggs

1. Escherichia coli is one of the most common facultative anaerobic species present in the gastrointestinal tract of animals and human beings. Usually they occur as commensals, but some serotypes can cause significant illnesses in humans as well as mammals and birds.

2. The occurrence of E. coli in different categories of table eggs collected from markets was evaluated. Isolates were analysed for the presence of virulence genes, antibiotic susceptibility pattern and efficacy of peracetic acid and chlorine for the purpose of decontaminating table eggs.

3. Significant differences were observed in the occurrence of E. coli between different groups viz. processed (cleaned, washed, sanitised and packed eggs), unprocessed (un-cleaned, un-sanitised and loose eggs) and free range (eggs obtained from backyard poultry) table eggs. Overall, E. coli occurred in table eggs at 28.6% with 22.9, 29.2 and 50.0% occurrence in processed, unprocessed and free-range table eggs, respectively.

4. A total of 24 isolates of E. coli were obtained and screened for virulence genes viz. STH, SLT1/2 and INVE genes. Of the 24 isolates recovered, 10 typeable isolates belonged to O141, O119, O9, O120 and O101 serotypes, while the remaining 14 were untypeable. Antibiograms of the isolates showed multiple antimicrobial resistance (MAR) index in the range of 0.13–0.40.

5. Peracetic acid (PAA) and chlorine (CL) were studied for their sanitisation efficacy; concentrations of 100 mg/kg of PAA and 200 mg/kg of CL completely inactivated E. coli over the egg surface and also resulted in 2.58 and 2.38 log reduction in total viable counts (TVC), respectively.

6. The presence of virulence-associated shiga-like toxin (SLT1/2) and invasion E (INVE) genes and antimicrobial resistance among the emerging serotypes of pathogenic E. coli isolated from table eggs has public health implications. It underscores the need to implement better management practices across the production systems and marketing channels to produce E. coli-free wholesome eggs for consumers.     

The effect of keel fractures on egg production,feed and water consumption in individual laying hens

1. The impact of keel bone fractures on egg production, egg weight and feed and water consumption in individual laying hens. A total of 165 Lohmann brown laying hens were obtained from a commercial farm that consisted of 105 with keel fractures and 60 without keel fractures.

2. After a 4-d period of acclimatisation, hens were individually housed and provided with ad libitum food and water for a 24-h period. The number of eggs laid, egg weight, feed and water consumption during this period were recorded. Keel bone strength was also assessed.

3. Hens free from keel fractures laid more eggs (91.7% vs. 84.9%) of significantly heavier weight (61.9 g vs. 60.2 g), ate less feed (139 g vs. 151 g) and drank less water (212 ml vs. 237 ml) than hens with fractures.

4. There was a significant positive association between keel fracture severity and water consumption, and a significant negative association between keel fracture severity and egg weight and keel bone strength.

5. This small-scale study on individual birds shows that keel bone fractures may have an impact on the economics of egg production.     

Comparisons of egg quality traits,egg weight loss and hatchability between striped and normal duck eggs

1. The egg quality of striped and normal duck eggs was compared to determine why striped eggs show decreased hatchability. A total of 430 eggs, obtained from a Pekin duck breeder flock aged 50–65 wks, were used in three experiments. The eggs were weighed and assigned randomly to measure egg quality traits, egg weight (EW) loss and hatchability during incubation.

2. There were no significant differences between egg types in terms of egg shape index, eggshell strength and thickness, albumen height, Haugh unit, yolk colour, weight of the eggshell with or without membranes, calcium, phosphorus, copper and manganese contents in the eggshell (with the inner and outer membranes or without the inner membrane), albumen weight, dry matter of albumen, crude protein (CP) of thick albumen and pH of the thick albumen.

3. The weight of eggshells with membranes, weight of thick albumen and CP of thin albumen in striped eggs were lower than those in normal eggs.

4. The thin albumen in striped eggs was heavier than that in normal eggs. The pH of the thin albumin in striped egg was significantly higher than that in normal eggs.

5. There were no significant differences in EW loss during incubation or duckling weight between striped and normal eggs. However, the hatchability of striped eggs was lower.

6. The lower weight of the eggshell inner membrane and thick albumen, lower CP content and higher pH in the thin albumen of striped eggs might contribute to lower hatchability.     

Enrichment of conjugated linoleic acid (CLA) in hen eggs and broiler chickens meat by lactic acid bacteria

1. The aim of this work was to compare conjugated linoleic acid (CLA) concentrations in chickens supplemented with 4 American Tissue Culture Collection (ATCC) bacterial strains, Lactobacillus plantarum, Lactobacillus lactis, Lactobacillus casei and Lactobacillus fermentum, and 4 isolates of Lactobacillus reuteri from camel, cattle, sheep and goat rumen extracts.

2. Micro-organisms were grown anaerobically in MRS broth, and 10⁶ CFU/ml of bacteria were administered orally to mixed-sex, 1-d-old broiler chickens weekly for 4 weeks and to 23-week-old layer hens weekly for 6 weeks.

3. The 4 strains were evaluated for their effects on synthesis of CLA in hen eggs and broiler meat cuts.

4. Administration of pure Lactobacillus and isolated L. reuteri strains from camel, cattle, goat and sheep led to significantly increased CLA concentrations of 0.2–1.2 mg/g of fat in eggs and 0.3–1.88 mg/g of fat in broiler chicken flesh homogenates of leg, thigh and breast.

5. These data demonstrate that lactic acid bacteria of animal origin (L. reuteri) significantly enhanced CLA synthesis in both eggs and broiler meat cuts.     

Defective egg production in a population of dwarf white leghorns 1

1. The incidence of defective eggs during the first 60 weeks of age increased with time due to significant increases in broken and membranous eggs in a population of dwarf White Leghorn hens.

2. The production of double‐yolked eggs/100 hen d, high during the initial weeks of lay, rapidly declined and disappeared by 30 weeks of age.

3. The frequencies of broken, membranous and soft‐shelled eggs were significantly correlated with each other and with the frequency of multiple daily ovulations indicating a common aetiology for these defective egg types.

4. Double‐yolked eggs were produced by the heavier, earlier‐maturing pullets in the population.

5. Normal egg production in dwarf White Leghorns may be enhanced by reducing the incidence of defective eggs.     

Comparative study of the effects of organic selenium on hen performance and productivity of broiler breeders

1. A 10-week experiment was conducted with Ross 308 broiler breeder chickens in cages to evaluate the influence of organic and inorganic sources of selenium (Se) supplementation. A total of 600 birds at 29 weeks of age were divided at random into 4 groups and fed on a maize–soya basal diet supplemented with different forms of Se.

2. The first (control) group was given the basal diet without Se supplementation, whereas the second, third and fourth groups were given, respectively, the basal diet with 0.3 mg/kg of inorganic Se in the form of sodium selenite or 0.3 and 0.5 mg/kg of organic Se in the form of Se enriched yeast (Se-yeast).

3. The experiment was carried out for 10 weeks to compare and evaluate the influence of Se supplementation on breeder performance, egg production, hatchability and the quality of eggs. Samples were collected for analysis at weeks 0, 5 and 10 of the experimental period.

4. At the end of the experiment (39 weeks), there was a reduction in mortality in breeders given diets supplemented with 0.5 mg/kg of Se-yeast. Supplementation of feed with 0.5 mg/kg organic Se increased egg production, percentage of settable eggs and hatchability.

5. Selenium supplementation increased egg weight and specific gravity compared to the control diet. However, no significant variation was found in albumen or yolk protein content at the end of week 10.

6. Selenium accumulation and glutathione peroxidase (GSHPx) activity were lower in the egg albumen and yolk of control compared with Se-supplemented treatments. Se accumulation and GSHPx activity were higher in the group given 0.3 mg/kg organic compared to 0.3 mg/kg of inorganic Se.

7. The results favour the use of Se-yeast at the dose of 0.5 mg/kg in broiler breeder diets for better productivity of eggs, settable eggs, hatchability and higher Se accumulation and antioxidant status in eggs.     

Embryonic development in the ostrich (Struthio camelus) during the first 7 days of artificial incubation

1. Early development of ostrich embryos was investigated in relation to time of egg collection and genotype.

2. A total of 321 ostrich eggs were collected during the 2008 and 2009 breeding seasons and the development of the embryo for up to the first 168 h of incubation was described and analysed. A sample of the incubated eggs was weighed and opened daily to investigate developmental changes.

3. In fresh eggs, the blastoderm contained a round, translucent dark area pellucida (AP) in the centre, with a surrounding thin white ring, likely to be the beginning of the area opaca (AO). Fresh eggs were considered infertile if the blastoderm was absent and instead numerous white droplets were present surrounded by vacuoles.

4. The average blastoderm area of a fresh fertile egg was 15.8 mm², increasing to 143.3 mm² after 2 d of incubation. By 72 h of incubation the area vasculosa (AV) was discernible in the posterior half of the blastoderm.

5. At 48 h of incubation the blastoderm area in eggs from the South African Black genotype (SAB) × Zimbabwean Blue genotype (ZB) crosses (104.5 ± 18.6 mm²) was lower than the pure SAB (141.0 ± 10.5 mm²), ZB (161.7 ± 13.5 mm²) and ZB × SAB crosses (166.1 ± 14.2 mm²).

6. Embryo length was 5.01 mm after 72 h of incubation and 14.5 mm after 168 h of incubation. At 168 h of incubation AV lengths for both ZB × SAB (53.2 mm) and SAB × ZB crosses (54.1 mm) were longer than in embryos from the pure breeds.

7. Results from this study can be put to practical use when determining whether eggs are infertile or fertile and also in investigating the age of early embryonic mortalities.     

Effect of egg composition on hatchability and on growth and slaughter characteristics of meat-type chicks

1.?Using the so-called TOBEC (Total Body Electrical Conductivity) method, which allows the determination of egg composition in vivo, correlations between egg composition, hatchability and hatched chicks’ development were studied.

2.?A total of 1500 hen eggs (Ross-308) were measured by TOBEC, and eggs with extremely high (10%, n = 150), extremely low (10%, n = 150) and average (10%, n = 150) electrical conductivity values were chosen for further investigation.

3.?During the incubation period, it was observed that eggs with high electrical conductivity had a significantly higher mortality than eggs with low electrical conductivity.

4.?It was observed that both the increase in electrical conductivity at the same egg weight, and the increase in egg weight at the same electrical conductivity resulted in an increase in the hatching weight.

5.?It was found that the dry matter, protein and fat content of the chicks hatched from eggs with low electrical conductivity was higher at hatching than that of the chicks hatched from eggs with high electrical conductivity.

6.?At 42 d of age the liveweight of cocks and pullets hatched from eggs with low electrical conductivity was 3·2 and 8·2% higher than the liveweight of cocks and pullets hatched from eggs with high electrical conductivity.

7.?Because of the higher liveweight at slaughter, there was a significant superiority of the chicks hatched from eggs with low electrical conductivity in the case of the examined carcase traits at slaughter.

8.?Similar tendencies were found also in the ratios of carcase variables to liveweight, but the between group differences were not statistically significant in this case.

9.?Based on the results it was concluded that TOBEC seems to be a useful method for separating eggs with different composition.

10.?This could be a good starting point for further in vivo investigations in order to clarify the effect of egg composition on hatchability and further development.     

Effects of feeding programme and feeder space change at photo-stimulation using maize- or wheat-based diet on growth and reproductive performance of female broiler breeders

1. This study was carried out to determine the effects of breeder feeding programme to 29 weeks of age, and feeder space change at photo-stimulation using two sources of grain on breeder hen reproductive efficiency and egg characteristics.

2. Fast-feathering Cobb 500 pullet broiler breeders were housed in 16 pens of 81 females each during rearing, and fed on either maize- or wheat-based diet, formulated to have similar nutrient composition. Two feeding programmes, Fast and Slow, were used from 14 to 29 weeks of age. At 22 weeks of age, 69 females that represented the body weight (BW) distribution from each pen were placed in a layer house where feeder space either remained very similar (from 6.3 to 6.5 cm/female) or was increased (from 6.3 to 8.4 cm/female). Breeder growth performance, reproductive efficiency and egg characteristics were evaluated.

3. Data were analysed as a 2 × 2 × 2 factorial design with diet type, feeding programme and feeder space change as the main factors. The main effects of the treatments were found but there were no treatment interactions.

4. Breeders fed on wheat were consistently heavier than breeders fed on maize from 10 to 52 weeks of age and exhibited greater hen mortality during the layer phase. Breeders fed on wheat diets exhibited improved egg production while laying eggs with a greater yolk:albumen ratio compared to the maize group.

5. The Fast feeding programme increased female mortality and increased BW during the layer phase. Breeders fed according to the Slow feeding programme had better fertility, and laid eggs with lower percentage eggshell that might be associated with the lower early embryonic mortality observed in these hens as compared with the Fast feeding programme.

6. Breeders having increased feeder space at photo-stimulation matured earlier and produced more eggs compared to breeders with no change in feeder space.

7. It was concluded that the feeding of wheat, the use of the Slow feeding programme and an increase in feeder space improved some aspects of hen productivity and egg characteristics.     

Flavobacterium psychrophilum Infections in Salmonid Broodstock and Hatchery-Propagated Stocks of the Great Lakes Basin

Bacterial coldwater disease (BCWD), caused by Flavobacterium psychrophilum, threatens wild and propagated salmonids worldwide and leads to substantial economic losses. In addition to being horizontally transmitted, F. psychrophilum can be passed from infected parents to their progeny, furthering the negative impacts of this pathogen. In Michigan, both feral and captive salmonid broodstocks are the gamete sources used in fishery propagation efforts. A 5-year study was initiated to follow the prevalence of systemic F. psychrophilum infections in feral broodstocks of four species (steelhead Oncorhynchus mykiss [potadromous Rainbow Trout]; Coho Salmon O. kisutch; Chinook Salmon O. tshawytscha; and Atlantic Salmon Salmo salar) residing in three Great Lakes watersheds. Additionally, captive broodstocks of four species (Rainbow Trout, Brown Trout Salmo trutta, Lake Trout Salvelinus namaycush, and Brook Trout Salvelinus fontinalis) maintained at two facilities were assessed for the presence of F. psychrophilum. The resultant offspring from each broodstock population were sampled for F. psychrophilum infections multiple times throughout hatchery residency. Using selective flavobacterial culture and PCR confirmation, F. psychrophilum was detected in all broodstocks except the captive Lake Trout and Brook Trout. Logistic regression analysis demonstrated that among the infected feral broodstocks, Chinook Salmon from the Lake Michigan watershed had the highest prevalence of systemic F. psychrophilum infection (mean = 63.2%). Among the captive broodstocks, the Gilchrist Creek strain of Brown Trout had the highest infection prevalence (mean = 5%). Collectively, the captive broodstocks were found to have significantly lower infection prevalence than the feral broodstocks. Despite the high prevalence of systemic F. psychrophilum infections in many broodstock populations, the bacterium was rarely detected in their progeny during hatchery rearing. However, heavy losses associated with clinical BCWD outbreaks did occur. Collectively, our results reinforce that BCWD continues to threaten Great Lakes basin salmonids.

Received April 6, 2015; accepted August 25, 2015     

Influence of citrus flavonoids on laying hen performance,inflammatory immune response,egg quality and yolk oxidative stability

1. The objective of this study was to investigate the effects of dietary supplementation with natural flavonoids (naringin and hesperidin) on laying hens’ performance, cellular immunity and egg quality parameters.

2. A total of 72 individually caged laying hens were allocated into 1 of 6 treatment groups: a control (C) group that was fed with a basal diet and groups that were offered the same diet further supplemented with hesperidin at 0.75 g/kg (E1), or 1.5 g/kg (E2), or naringin at 0.75 g/kg (N1), or 1.5 g/kg (N2) or α-tocopheryl acetate at 0.2 mg/kg (VE) for 67 d.

3. Supplementation with naringin or hesperidin did not affect the performance and egg quality (P > 0.05) apart from an improvement in the yolk colour that was more orange in naringin and hesperidin groups in comparison to the controls (P-linear < 0.05). Egg yolk and plasma cholesterol levels were not affected by citrus flavonoids (P-linear > 0.05).

4. Inflammatory immune response, measured by phytohaemagglutinin skin test (PHA), was suppressed in laying hens that were fed with either naringin (P-linear < 0.05) or hesperidin (P < 0.05). Egg yolk oxidative stability was improved from the 4th d after naringin or hesperidin supplementation. This beneficial effect was comparable to that of α-tocopheryl acetate and was observed in eggs that were stored for up to 120 d.

5. In conclusion, naringin and hesperidin may favourably prolong the shelf life of eggs, appear to possess anti-inflammatory properties and could improve the yolk colour without any side effects on the performance or egg quality traits.     

Physical characteristics of intact and cracked eggs

1. Six egg samples, comprising equal numbers of intact and cracked eggs, were collected from three flocks of hybrid layers.

2. Cracks were classified according to type (hole, star or straight) and the weight, shape, shell colour and specific gravity of all the eggs were measured. Shell thickness and dry shell weight were also recorded for a maximum of 10 intact and 10 cracked eggs from each sample.

3. The largest proportion of cracks were holes.

4. Differences in mean egg weight and shape of intact and cracked eggs within each sample were inconsistent. Mean shell colour was darker for the intact brown eggs than for the cracked ones. For all the samples, mean egg specific gravity, shell thickness and dry shell weight were higher for the intact eggs. The difference in egg specific gravity was significant (P<0.05) for all samples.

5. Holes occurred in eggs of lower mean weight but with better quality shells than those in which star or straight cracks occurred.     

Effects of dietary supplementation with Gynura procumbens (Merr.) on egg yolk cholesterol,excreta microflora and laying hen performance

1. The present study investigated the effects of dietary supplementation with Gynura procumbens on egg yolk and serum cholesterol and triglycerides, excreta microflora, laying performance and egg quality.

2. A total of 160 Hy-Line Brown layers (45 weeks old) were randomly assigned into 4 treatments on the basis of laying performance. Each treatment had 4 replicates with 10 birds each.

3. Dietary treatments were basal diet supplemented with 0 (control), 2.5, 5.0 and 7.5 g/kg diet G. procumbens during 56-d feeding period.

4. Serum (d 21, 42 and 56) and egg yolk (d 28, 42 and 56) cholesterol and triglycerides concentrations were linearly reduced with increasing dietary concentrations of G. procumbens.

5. Increasing dietary concentrations of G. procumbens linearly reduced the excreta total anaerobic bacteria (d 28), Clostridium sp. and Escherichia coli (d 28 and 56) populations.

6. Overall egg production and egg mass were linearly increased, and overall feed efficiency was linearly improved with increase in dietary G. procumbens.

7. Dietary increasing concentrations of G. procumbens linearly improved egg yolk colour (d 28 and 56) and breaking strength of eggs (d 56).

8. The results obtained in the present experiment indicate that dietary supplementation with G. procumbens could reduce the egg yolk cholesterol, suppresses harmful excreta microflora and improves layers performance.     

Depletion of florfenicol and florfenicol amine residues in chicken eggs

1. The aim of this study was to develop a suitable method for the analysis of florfenicol (FF) and its metabolite florfenicol amine (FFA) in chicken eggs and to determine FF and FFA residue depletion in eggs of laying hens.

2. The analytes were extracted from yolk, albumen and whole egg by phosphate buffer and ethyl acetate. Following purification, samples were analysed by high-performance liquid chromatography.

3. Fifty laying hens were divided into 5 groups, and each hen received doses of 20 mg/kg FF: Group 1 (received a single oral dose by gavage); Group 2 (a single intramuscular dose); Group 3 (a single subcutaneous dose); Group 4 (multiple oral doses for 3 d) and Group 5 (multiple oral doses for 5 d).

4. Limits of detection and of quantitation values were 1.94 and 6.45 g/10⁹ g (ppb) for FF, respectively, and 0.48 and 1.58 ppb for FFA, respectively. Relative standard deviation values of intra-day and inter-day variation below 11% also confirmed the usefulness of the method for analysing FF and FFA in eggs.

5. From the first day of both oral and parenteral administration, FF and FFA were detected at 0.1% and 0.08% of dosage, respectively, and 57% of the drugs were eliminated from the egg yolk. Elimination time of FF was 8 d in Groups 1, 2 and 3; 9 d in Group 4 and 10 d in Group 5.