Respiratory phagocytes are implicated in enhanced colibacillosis in chickens co-infected with influenza virus H9N2 and Escherichia coli

1. The aim of this study was to determine the most likely time interval after infection with influenza virus H9N2 for co-infection with Escherichia coli to cause colibacillosis, the importance of lung load of E. coli and the involvement of respiratory phagocytes.

2. Specific pathogen free chickens were inoculated intranasally with 10⁶EID₅₀ of influenza virus or uninfected. After specified time intervals, 10⁷ CFU E. coli or phosphate-buffered saline was inoculated. The presence of lesions, the number of respiratory phagocytes in the respiratory lavage fluid and the E. coli load in the lung were determined after different time intervals.

3. Compared with the number of lesions in chickens receiving only E. coli inoculation, the number lesions in co-infected chickens were increased at 0- and 3-d time intervals, but reduced in the groups at 6- and 9-d intervals between co-infection.

4. At 1–3 d after E. coli inoculation, the number of lesions chickens was correlated with the number of respiratory phagocytes harvested and related to the E. coli load in the lungs at 5 d.

5. These results suggest that the lesions caused by E. coli in chickens were increased within a 0–3 d interval following H9N2 virus inoculation and that this effect is related to the number of respiratory phagocytes.     

Molecular characterisation of Escherichia coli from dead broiler chickens with signs of colibacillosis and ready-to-market chicken meat in the West Bank

1. The aim of this work was to compare a group of virulence-associated characteristics of Escherichia coli isolates from broiler chickens that had died with signs of colibacillosis against E. coli isolates from ready-to-market chicken meat in the West Bank.

2. The isolates were investigated to determine the virulence factor (VF) profile, phylogenetic group and the presence of extended-spectrum beta-lactamase (ESBL). A total of 66 avian pathogenic E. coli (APEC) strains from different affected broiler farms and 21 E. coli isolates from ready-to-market chicken carcasses (hereinafter called meat strains) from 8 slaughter houses were analysed.

3. The overall content of VFs was significantly higher (P < 0.05) among APEC strains, with over 75% of APEC strains having ≥4 VFs, while over 75% of the meat strains had <4 VFs. The VFs iss, astA and iucD were frequently detected in APEC and meat strains, whereas cvi, papC, vat, tsh and irp2 occurred more significantly in APEC strains. Phylogenetic typing showed that 67% of the meat strains belonged to group B2. Phylogroup D was predominant (50%) in the APEC strains. Using double disc diffusion and polymerase chain reaction (PCR), 10.6% of the APEC and 9.5% of the meat strains were determined to be ESBL positive.

4. Our findings show that the VFs papC, vat, irp2 and to a lesser extent tsh and cvi are significantly more prevalent in APEC strains. The results demonstrate that chicken meat can be contaminated with APEC strains (≥4 VF). A significant percentage of the meat strains fall in the B2 group, which is a phylogroup largely associated with human pathogenic ExPEC strains. The results of ESBL screening indicated that broiler chicken products in Palestine represent a potential reservoir of ESBL genes and therefore could be considered a possible public health risk.     

The presence of salmonellae during the rearing of Turkeys

1. The incidence of salmonellae was investigated in two batches of turkeys in the environmentally‐controlled houses and naturally ventilated pole‐barns of a commercial turkey enterprise.

2. The poults were free from contamination on arrival at the rearing house but were contaminated via the water supply from troughs which had been ineffectively fumigated.

3. Foodstuffs did not appear to be a source of contamination.

4. The spread of salmonellae in this turkey rearing enterprise was very similar to that in chicken rearing houses.     

The association between extended spectrum beta-lactamase (ESBL) and ampicillin C (AmpC) beta-lactamase genes with multidrug resistance in Escherichia coli isolates recovered from turkeys in Brazil

1. The aim of this study was to analyse the association between Escherichia coli isolates recovered from turkeys and the expression of beta-lactamase genes, such as extended spectrum beta-lactamase (ESBL) and ampicillin class C (AmpC). The phenotype of the resistance profile was examined using the association between amoxicillin and ceftiofur resistance.

2. Results showed that 84% from the turkey isolates harboured 4 or 5 genes associated with the CoIV plasmid. In an antibiogram test, 82% of the isolates were multidrug-resistant, the highest levels of resistance being against erythromycin (99%) and amoxicillin (76.1%). ESBL-positive groups were 31% positive for the ctx-m-2 gene, 6.8% were positive for ctx-m-8 and 70% harboured the tem wild gene.

3. All positive isolates from the AmpC beta-lactamase-positive group harboured the cmy-2 gene. The presence of the cmy-2 gene was associated with both the CTX-group genes and resistance to ceftiofur.

4. There was a high prevalence of avian pathogenic E. coli in suspected cases of colibacillosis in turkeys and a high antimicrobial resistance index. The results highlighted the risk of ceftiofur resistance and the presence of both ESBL and AmpC beta-lactamase E. coli in the turkey production chain.     

Immune-modulatory effects of dietary Saccharomyces cerevisiae cell wall in broiler chickens inoculated with Escherichia coli lipopolysaccharide

1. An experiment was conducted to study the effect of yeast (Saccharomyces cerevisiae) cell wall (YCW) supplemented in diets of broiler chickens challenged with Escherichia coli lipopolysaccharide (LPS).

2. One-day-old broiler chicks were randomly distributed into 24 cages (6 replicate cages; 8 birds/cage) and were inoculated with 0 or 1 mg/kg body weight E. coli-LPS (d 4 and 9) and 0 or 500 mg YCW/kg feed, resulting in a 2 × 2 factorial arrangement of treatments. Experimental diets did not include coccidiostats, in-feed antibiotics or enzymes.

3. On d 21, the inoculation of E. coli-LPS reduced weight gain and feed intake and increased feed conversion ratio (FCR) of birds, an effect maintained until 28 d. In contrast, chickens given diets with YCW improved the FCR at both 21 and 28 d of age.

4. E. coli-LPS challenge reduced the relative weight of bursa of Fabricius, except when chickens were given YCW, which resulted in an interaction. Supplementation of broiler diets with YCW exacerbated the cellular immune response as measured by the delayed cutaneous hypersensitivity response test.

5. The results of this study suggested a benefit on feed efficiency when YCW was added to diets fed to broiler chickens challenged with E. coli-LPS. Part of the mode of action of YCW might be related to better maintenance of immune status in response to microbial challenge.     

Performance,gut morphology and microbiology effects of a Bacillus probiotic,avilamycin and their combination in mixed grain broiler diets

1. This study aimed to determine the effect of avilamycin (AGP) and a multi-strain Bacillus probiotic (DFM) on the performance, gut histology and microbiology of broilers fed on a mixed grain diet.

2. A total of 800 chicks were allocated to four treatments: a control diet, control+AGP, control+DFM, or control+AGP+DFM. Bodyweight, feed intake and FCR were measured at d 0, 21 and 42. Samples were taken at d42 to determine villus height (VH), crypt depth (CD) and ratio (VH:CD). Mucosal E. coli and Lactobacilli counts were measured at d42.

3. At d42, DFM and AGP+DFM significantly increased weight over the control, with AGP returning an intermediate value. FCR followed a similar pattern. DFM and AGP+DFM significantly increased VH and CD in all gut sections compared to the control.

4. DFM and AGP+DFM reduced E. coli counts compared to control, with AGP reducing caecal counts only, while Lactobacilli counts were increased.

5. Divergent histology and microbiology between treatments highlight the different modes of action of AGP and DFM for improving broiler growth and feed efficiency.     

Volatile gas concentrations in turkey houses estimated by Fourier Transform Infrared Spectroscopy (FTIR)

1. The aim of the present study was to estimate gas concentrations in commercial turkey houses by Fourier Transform Infrared Spectroscopy (FTIR).

2. The experiment was conducted in 5 buildings of a commercial turkey farm. The measurements of gases were carried out every 3 weeks of the growth cycle.

3. The results demonstrate that ammonia and carbon dioxide are the prevalent gases released during the entire production cycle in turkey houses. The mean concentrations of the above compounds ranged between 4–31 ppm and 220–2058 ppm, respectively. Thiols, nitriles, amines, aldehydes, hydrocarbons and other organic and inorganic compounds also occurred in turkey houses, but they were emitted periodically and their mean concentrations were significantly lower in comparison with CO₂ and NH₃.

4. Lower ventilation ratio and higher moisture of excreta in the first half of the growth period accelerated the release of some gases, whereas gradual faeces and urine accumulation contributed to an increase in the concentration of selected organic compounds.

5. A portable FTIR analyser is a useful device for measuring gas concentrations in commercial turkey farms, and it supports determinations of tolerable emission limits in turkey production.     

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.     

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.     

Effect of dietary supplementation of bacteriophage on performance,egg quality and caecal bacterial populations in laying hens

1. Bacteriophages (BP) have gained increasing attention as a treatment of bacterial infection for animals. However, the data pertaining to dietary application of BP for laying hens have been limited.

2. This study aimed to investigate the effect of dietary BP on laying performance, egg quality and caecal bacterial populations in laying hens.

3. The dietary BP used in this experiment was a mixture of individual BP targeting Salmonella gallinarum, Salmonella pullorum, Salmonella typhimurium, Salmonella enteritidis, Salmonella derby and Staphylococcus aureus.

4. A total of 360 Hy-Line Brown laying hens of 32 weeks of age were allotted to one of three dietary treatments with 6 replicates in a completely randomised design. The basal diet was prepared, and 0.4 or 0.8 g/kg BP mixture was supplemented to the basal diet. Diets were fed to hens for 8 weeks.

5. Laying performance and egg quality were not affected by dietary treatments. As inclusion levels of BP mixture in diets were increased, the DNA copy numbers for Salmonella spp. in the caecal contents decreased linearly, whereas the DNA copy numbers for Escherichia coli in the caecal contents increased linearly.

6. Results indicate that dietary supplementation of BP mixture decreases the target Salmonella spp. populations but increases Escherichia coli populations in the gastrointestinal tract of laying hens with little impact on laying performance and egg quality.     

The effect of continuous aeration on the fertility of fowl and Turkey semen stored above 0 °C

1. Fowl and turkey spermatozoa were found to utilise oxygen when stored in tubes at 5 and 10 °G respectively; the storage medium becoming rapidly anaerobic.

2. This aerobic metabolism was encouraged by continuous introduction of air into the semen during storage.

3. Fertility of turkey semen stored at 10 °C for 24 h in a diluent containing glucose was increased 24‐fold by simple aeration and reached 99% of that achieved with fresh semen.

4. Fertilities of greater than 90% were also achieved with fowl semen stored aerated for 48 h at 5 °C in a diluent with or without glucose. The increased fertility on aeration was greater (2.2‐fold) with fowl semen stored in the absence of glucose.     

Prevalence of tetracycline resistance determinants in broiler isolated Escherichia coli in Iran

1. Tetracycline resistance determinants are widespread among bacterial species. Resistance to tetracycline occurs by different mechanisms regulated by various genes.

2. The study was conducted to determine the tetracycline resistance and prevalence of tetracycline resistance determinants among Escherichia coli strains isolated from broilers in northern Iran.

3. Minimum inhibitory concentration (MIC) of tetracycline and susceptibility pattern of the isolates were screened using micro-dilution and disk diffusion methods, respectively. The presence of 7 tetracycline resistance genes including tetA, tetB, tetC, tetD, tetE, tetG and tetM was tested using the polymerase chain reaction.

4. Among 100 strains isolated from broilers, 73% were identified as tetracycline resistant. All isolates showed the presence of tetracycline-associated genes. The most prevalent genes were tetA (46%) and tetB (41%) and totally, 17 different genotypes were recognised according to the presence of tetracycline resistance genes. Statistical analysis revealed that concomitant presence of the resistance genes significantly increased the tetracycline MIC and effectiveness of phenotypic characterisation.

5. The results demonstrated a high occurrence of tetracycline-resistant E. coli and related genes among broilers which presents a risk of increasing these strains in human infections associated with food animals.     

Concentrations of plasma luteinising hormone,prolactin, progesterone and androgens during the ovulatory cycle of the Turkey

1. Changes in the concentrations of plasma luteinising hormone (LH), prolactin, androgen and progesterone were measured during the ovulatory cycle of the turkey.

2. Single pre‐ovulatory peaks of plasma LH, androgen and progesterone were observed which took 8, 8 and 12 h respectively, to increase and return to base‐line values. The concentration of plasma prolactin tended to be elevated between 6 h before and 6 h after the LH peak with the maximum values occurring after the peak.

3. The changes in the concentrations of plasma LH and progesterone were 3‐ and 7‐fold respectively while 2‐fold changes were observed in the concentrations of plasma androgen and prolactin.

4. The pre‐ovulatory concentration of plasma progesterone and prolactin began to decrease 4 and 6 h respectively, after the pre‐ovulatory peak of LH.

5. Ovulation and oviposition occurred 6 to 8 h and 36.10+ 0.57 h (SEM) ( n= 11) respectively after the pre‐ovulatory peak of LH.

6. In birds kept on 14 h light/d, pre‐ovulatory peaks of LH were initiated only during a 10 to 11‐h period starting within 2 h after the onset of darkness.

7. A comparison between these data and those from the fowl suggest that the egg is retained in the turkey's oviduct for about 3 to 4 h longer than in the fowl.     

Encapsulated benzoic acid supplementation in broiler diets influences gut bacterial composition and activity

1. The present study investigated the effects of encapsulated benzoic acid (BA) supplementation in broiler feed on performance and gastrointestinal microbiota.

2. Eighty broilers were randomly divided into two groups. Birds in the control group were fed on maize–soybean-based diets. Birds in the treatment group were provided the same diet supplemented with 2 g/kg BA encapsulated in a vegetable oil matrix.

3. At the end of the trial (d 35), pH, bacterial composition and metabolites were determined in the crop, jejunum, ileum and caecum.

4. Growth performance variables and pH were not significantly different.

5. BA concentration decreased rapidly in the proximal gut. However, the treatment diet showed higher BA in the crop, jejunum, ileum and caecum.

6. Total lactate in the crop and D-lactate in the jejunum was higher in the BA treated group. Caecal total and branched chain fatty acids were decreased due to the treatment.

7. Lactobacilli populations were significantly altered by BA supplementation. A trend for increased lactobacilli was observed in the crop, while it became significant in the jejunum and ileum. Lactobacillus species responded differently to the treatment. Four of 5 measured Lactobacillus species, particularly in the ileum, followed the course observed for total lactobacilli; only Lactobacillus salivarius was not modified.

8. Correlation analysis showed that BA modified the intestinal microbiota. Lactobacilli correlated negatively to all studied clostridial clusters and enterobacteria. Clostridial clusters IV and XIVa were significantly increased in the jejunum, whereas only clostridial cluster XIVa was increased in the caecum.

9. Encapsulated BA modified the intestinal microbiota which can lead to the conclusion, that the main beneficial mode of action of BA in the gut appears to be the enhancement of lactic acid bacteria, which in turn may act as a vanguard against pathogens.     

A mathematical representation of the Ovulatory cycle of the domestic hen

1. The ovulatory cycle of the domestic hen (Gallus domesticus) was postulated to be the result of the interaction of two independent systems.

2. A circadian system was postulated to control the restriction of ovulation to an 8‐h period of the day under conventional 14 h light: 10 h dark regimes.

3. The final phase of follicular maturation was postulated to commence after ovulation of the preceding ovum in the hierarchy.

4. Ovulation was postulated to occur when a mature follicle was present in the ovary during the appropriate phase of the circadian‐linked system.

5. The predicted times of oviposition were within the standard error of the observed times of oviposition under 21‐, 24‐ and 28‐h photoschedules.

6. It was concluded that this hypothesis for the control of the ovulatory cycle of the hen is consistent with current knowledge.     

The effects of restricted feeding on adrenal cortical activity in the immature domestic fowl

1. The effect of reducing food intake to 75% of the ad libitum intake was determined from hatching to 8 weeks in young Light Sussex chickens.

2. Restricted birds were lighter throughout the experiment.

3. Relative adrenal weight tended to be greater in restricted birds but the difference decreased with time.

4. There was no depletion of adrenal cholesterol: from week 5 there was a significantly greater amount in the adrenals of restricted birds.

5. After 1 week of restriction plasma corticosterone concentration was 73% greater than in controls. It decreased progressively, falling within the normal range at 5 weeks.

6. Restricted birds were hypoglycaemic from weeks 2 to 7 and hyper‐lipacidaemic throughout. A negative correlation between plasma glucose and free fatty acids was found.     

Susceptibility of various poultry species to dietary aflatoxin

1. The relative sensitivities of broiler chicks, White Leghorn chicks, quail chicks, goslings and turkey poults to increasing dietary contents of aflatoxin from 1 to 21 d of age were determined. Aflatoxin was included in the diets at three concentrations (0.7, 1.4 and 2.1 mg aflatoxin B_a equivalents /kg).

2. Data were analysed for species differences in food consumption, body weight, food utilisation, mortality and liver pathology.

3. There were no significant effects of aflatoxin in broiler and Leghorn chicks. Aflatoxin at 0.7 mg/kg decreased the body weight of turkey poults but tended to increase the growth rate of goslings. At 1 ‐4 mg aflatoxin/kg food consumption, body weight and weight gain of goslings were adversely affected.

4. Liver damage occurred in goslings and quail chicks at all inclusions of aflatoxin and was extensive at 2.1 mg/kg.

5. Poults and goslings appeared to be the most sensitive, quail were intermediate and domestic chicks were most resistant.     

Lipopolysaccharide reduces food passage rate from the crop by a prostaglandin-independent mechanism in chickens

1. We examined the effect of lipopolysaccharide (LPS), a component of Gram-negative bacteria, on food passage in the digestive tract of chickens (Gallus gallus) in order to clarify whether bacterial infection affects food passage in birds.

2. Food passage in the crop was significantly reduced by intraperitoneal (IP) injection of LPS while it did not affect the number of defecations, suggesting that LPS may affect food passage only in the upper digestive tract.

3. Similar to LPS, prostaglandin E2 (PGE2), one of the mediators of LPS, also reduced crop-emptying rate in chickens while it had no effect on the number of defecations.

4. Pretreatment with indomethacin, which is an inhibitor of cyclooxygenase (COX), a prostaglandin synthase, had no effect on LPS-induced inhibition of crop emptying.

5. IP injection of LPS did not affect the mRNA expression of COX2 in the upper digestive tract of chickens.

6. It is therefore likely that LPS and PGE2 reduced food passage rate in the crop by a prostaglandin-independent pathway in chickens.     

Effect of age and sex on the chemical composition of edible offal and blood from broilers

1. The effects of age and sex on the proximate composition and inorganic constituents in edible offal and whole blood from a commercial British broiler chicken strain were studied.

2. Although the protein content of all tissues varied with age, the only statistically significant effect was on the level of protein in the gizzard.

3. The moisture content of the gizzard was significantly affected by age and blood by age and sex.

4. The fat content of the gizzard was significantly affected by age, as was the effect of sex on the fat content of blood.

5. The ash contents of heart and gizzard were significantly affected by age.

6. The phosphorus contents of the gizzard, heart and liver were significantly affected by age, as were the chloride content of heart and the calcium content of the liver. Blood potassium was significantly affected by sex. Fluctuations in the contents of other elements are discussed.

7. The variability in organ composition is discussed with respect to the degree of trimming and preparation of the samples.     

Supplementary biotin decreases tibial bone weight,density and strength in riboflavin-deficient starter diets for turkey poults

1. Growth and skeletal responses to different dietary concentrations of riboflavin and biotin were compared in turkey poults from hatch to 21 d of age. The birds were fed on a turkey starter diet with different concentrations of supplementary riboflavin (0, 20 and 40 mg/kg) and biotin (0, 0.3 and 0.6 mg/kg) in a factorial design.

2. Poults fed on diets with no supplementary riboflavin had poor gait scores, decreased times to sit and higher rates of culling compared to poults fed on the control diet (20 mg biotin and 0.3 mg riboflavin/kg diet). Histologically, riboflavin deficiency was associated with a peripheral neuropathy similar to that described previously in chicks and, unexpectedly, in growth plate abnormalities.

3. Tibiae of poults fed on the control diet were larger, more dense, stronger and stiffer than the diets with no supplementary riboflavin.

4. Increasing supplementary biotin in poults fed on diets with no supplementary riboflavin was associated with a decrease in tibia weight, density, strength and stiffness.

5. The results demonstrated that riboflavin deficiency in fast-growing turkey poults was associated with growth retardation, growth plate disturbance and peripheral nerve dysfunction leading to an inability to walk.