Colorimetric Method of Loop-Mediated Isothermal Amplification with the Pre-Addition of Calcein for Detecting Flavobacterium columnare and its Assessment in Tilapia Farms

Flavobacterium columnare, the causative agent of columnaris disease in fish, affects many economically important freshwater fish species. A colorimetric method of loop-mediated isothermal amplification with the pre-addition of calcein (LAMP–calcein) was developed and used to detect the presence of F. columnare in farmed tilapia (Nile Tilapia Oreochromis niloticus and red tilapia [Nile Tilapia × Mozambique Tilapia O. mossambicus]) and rearing water. The detection method, based on a change in color from orange to green, could be performed within 45 min at 63°C. The method was highly specific, as it had no cross-detections with 14 other bacterial species, including other fish pathogens and two Flavobacterium species. The method has a minimum detection limit of 2.2 × 10² F. columnare CFU; thus, it is about 10 times more sensitive than conventional PCR. With this method, F. columnare was detected in gonad, gill, and blood samples from apparently healthy tilapia broodstock as well as in samples of fertilized eggs, newly hatched fry, and rearing water. The bacteria isolated from the blood were further characterized biochemically and found to be phenotypically identical to F. columnare. The amplified products from the LAMP–calcein method had 97% homology with the DNA sequence of F. columnare.

Received May 21, 2014; accepted August 10, 2014     

Evaluation of probiotics in diets with different nutrient densities on growth performance,blood characteristics,relative organ weight and breast meat characteristics in broilers

1. A total of 720 1-d-old broilers were used in a 28 d experiment to determine the effects of probiotic supplementation in diets with different dietary nutrient densities.

2. Birds were randomly allotted to one of the 4 treatments in a 2 × 2 factorial arrangement (12 replicateswith 15 broilers per replicate) with two levels of nutrient density [high nutrient density (metabolisable energy (ME) 12.7 MJ/kg and crude protein (CP) 230.3 g/kg for 1–7 d; ME 13.2 MJ/kg and CP 220.3 g/kg for 8–28 d) or low nutrient density (ME 12.1 MJ/kg and CP 220.2 g/kg for 0–7 d; ME 12.6 MJ/kg and CP 209.8 g/kg for 8–28 d)] and 0 or 2 g/kg probiotics (1.0 × 10¹⁰ viable spores/g of Bacillus subtilis endospores and 1.0 × 10⁹ viable spores/g of Clostridium butyricum).

3. The high-nutrient-density diet increased body weight gain (BWG), feed conversion ratio (FCR), serum cholesterol and triglyceride concentration relative to the low-nutrient-density diet. High-nutrient-density diet reduced water loss ratio of breast muscle, liver and fat relative to body weight compared to low-nutrient density-diet. The inclusion of probiotics increased BWG and feed intake throughout the experiment. Dietary probiotics increased the percentage of blood lymphocytes and relative weight of spleen and bursa of Fabricius when compared to the non-probiotic treatment. The inclusion of probiotics decreased serum cholesterol and triglyceride concentrations and lightness (L*) value of breast meat compared to the non-probiotic-supplemented diet.

4. In conclusion, high dietary nutrient density increased growth performance and serum cholesterol and triglyceride concentrations in broiler chickens. The inclusion of probiotics increased growth performance but reduced serum cholesterol and triglyceride concentrations. The positive effect of probiotic supplementation on growth performance was reduced by the high-nutrient-density diet during the first week of life.     

Growth performance and meat quality of broiler chickens supplemented with Rhodopseudomonas palustris in drinking water

1. The effect of the bacterium, Rhodopseudomonas palustris, on the growth performance and meat quality of broiler chickens was investigated.

2. A total of 900-d-old Arbor Acres broilers were allocated to three experimental treatments for 6 weeks. Chicks were administered with R. palustris in drinking water as follows: (i) control group without R. palustris; (ii) treatment 1 (R1) with R. palustris of 8 × 10⁹ cells per chick per day in drinking water; (iii) treatment 2 (R2) with R. palustris of 1.6 × 10¹⁰ cells per chick per day in drinking water.

3. The results showed that, compared with that of control, both groups of R. palustris treatment increased daily weight gain and improved feed conversion ratio of broiler chickens significantly during the whole growing period of 6 weeks.

4. Both total and glutamic acid contents of chicken breast fillet in R. palustris treatment R2 were higher, while the fat content was lower, than those of the control group. Furthermore, R. palustris treatments also improved sensory attributes of chicken breast fillet.

5. As a probiotic providing rich nutrients and biological active substances, R. palustris administration in drinking water displayed a growth promoting effect and improved meat quality of broiler chickens.     

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     

Pathogenicity of Members of the Vibrionaceae Family to Cultured Juvenile Sablefish

Sablefish Anoplopoma fimbria are a prized seafood species due to their high oil content and white flaky flesh. Raising these species in culture can help to provide an important source of protein for humans and relief to declining wild fish populations. Understanding the environmental factors that influence the production of Sablefish is important for successful culturing. The significance of host–pathogen interactions in Sablefish culture and the resulting environmental implications are unknown. Pathogens could potentially cause losses of cultured Sablefish stocks due to disease, while Sablefish cultured in net pens may also serve as reservoirs for pathogens and potentially transmit disease to wild fish species. In this initial study, the susceptibility of juvenile Sablefish to three bacterial pathogens from the family Vibrionaceae was examined. Listonella anguillarum, Vibrio ordalii, and V. splendidus can pose serious economic threats to cultured fish and shellfish. Groups of juvenile Sablefish were exposed to five concentrations of each of the pathogens. Sablefish were susceptible to L. anguillarum, but were resistant to V. ordalii and V. splendidus at exposure concentrations of ≤1.32 × 10⁷ CFU/mL and ≤3.57 × 10⁶ CFU/mL, respectively. The greatest L. anguillarum concentration examined (8.7 × 10⁶ CFU/mL) resulted in 24% mortality in juvenile Sablefish. A 24% loss of Sablefish stock could significantly influence an aquaculture program. As determined by multiple logistic regression, the survival of Sablefish to L. anguillarum exposure was significantly affected by their body mass, and larger fish had a greater probability of survival. Aquaculture operations could employ various strategies to minimize the loss of juvenile Sablefish by accounting for their size and known susceptibilities to pathogens.

Received December 9, 2014; accepted February 7, 2015     

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.     

Beneficial Effects of Rhodotorula sp. C11 on Growth and Disease Resistance of Juvenile Japanese Spiky Sea Cucumber Apostichopus japonicus

The purpose of this study was to evaluate the effects of dietary administration of the live yeast, Rhodotorula sp. C11, on growth and disease resistance against Vibrio splendidus infection in juvenile Japanese spiky sea cucumber Apostichopus japonicus. Sea cucumbers were fed diets containing Rhodotorula sp. C11 at 0 (control), 10⁴, 10⁵, and 10⁶ CFU/g of feed for 45 d. There were three replicate tanks per dietary treatment. The specific growth rates were higher in all sea cucumbers treated with Rhodotorula sp. C11 than in the controls. Following a challenge with V. splendidus NB13, the cumulative prevalence and mortality of sea cucumbers fed diets supplemented with Rhodotorula sp. C11 were lower than in animals fed the basal diet. In sea cucumbers fed diets supplemented with Rhodotorula sp. C11 for 42 d, the only viable yeast found in the intestine was Rhodotorula sp. C11, which had counts of 1.58–1.98 × 10⁴ CFU/g. No yeast was isolated from the intestine of animals fed the basal diet. For the colonization study, 20 sea cucumbers from each dietary treatment were removed to separate tanks and fed the control diet from day 16 to day 46. The viable yeast (Rhodotorula sp. C11) counts in the intestine decreased to 60–80 CFU/g by day 37. Moreover, as demonstrated by denaturing gradient gel electrophoresis, Rhodotorula sp. C11 colonization of the intestine could be detected until day 46. The differences in culture and PCR-denaturing gradient gel electrophoresis may be due to differences in the sensitivity of both methods. The present result showed that Rhodotorula sp. C11 was able to successfully colonize the intestine of juvenile Japanese spiky sea cucumbers by dietary supplementation, which improved its growth and disease resistance.

Received August 11, 2014; accepted November 14, 2014     

Thermoneutral zone and resting metabolic rate of broilers

1. Metabolic rate was determined once a week in broilers from a commercial source, from 1 to 63 d of age.

2. The equations relating minimal resting metabolic rate (oxygen consumption, ml/bird h,y) and body weight (W) were: males 45 to 497 g, y = 3.2 W^0.882 597 to 3000 g, y = 40.5 W^0.483; females 45 to 514 g, y = 2.52 W^0.881; 514 to 2500 g,y = 12.3 W^0.627.

3. The relationship between lower critical temperature (T_cl , °C,y) and age (d, x) may be described by the following equations: chicks 1 to 21 d, y = 34.2 ‐ 0.32 x; 14 to 63 d,y = 49.4 × ^?0194.

4. The relationship between T_cl and W may be described by one equation for both sexes between 100 and 3000 g, y = 62.15 W^?0.135..

5. The equations for T_cl and data for upper critical temperature (T_cu ) could be used to obtain maximal performance from broilers, with reduced costs, by providing a suitable environment related to age or body weight.     

Research note: methodology for high-quality RNA extraction from poultry whole blood for further gene expression analysis

1. There are no published methods for RNA isolation from avian whole blood where nucleated red blood cells prevent the use of established mammalian protocols. The aim of this study was therefore to develop a protocol for total RNA extraction using avian whole blood by defining the effect of anticoagulants and sample purification protocols on RNA yield and quality.

2. Blood collections from the cutaneous ulnar or medial metatarsal veins of birds yielded adequate blood volume (2–3 ml) draws. The experiment was a 2 × 2 × 3 factorial arrangement of treatments, with two levels of DNase (0 and TURBO DNA-free Kit), two levels of Cleanup (0 and RNeasy MinElute Cleanup Kit), and three anticoagulants (no anticoagulant, EDTA, or sodium citrate).

3. RNA was isolated successfully by adding TRIzol LS to 0.25 ml of chicken whole blood at 3:1 ratio. From 0.125 ml of avian whole blood, 2–3 µg of RNA with RNA integrity number values of 7.75 was successfully isolated with the TRIzol LS extraction and an RNeasy MinElute Cleanup Kit.

4. This reliable protocol can be used to extract high yield and quality of total RNA from a small amount of whole blood.     

Development of a PCR Assay Based on a Single–Base Pair Substitution for the Detection of Aeromonas caviae by Targeting the gyrB Gene

Aeromonas caviae is a bacterial pathogen that causes various infectious diseases in both humans and animals. To facilitate its detection, we developed species-specific primer sets targeting polymorphisms in the gyrB gene for use in a PCR assay. The technique was able to detect 100% (29/29) of the A. caviae strains tested using either of two sets of primers (designated ACF1-ACR and ACF3-ACR), which produced 293-bp and 206-bp amplicons, respectively. Another set of primers (designated ACF2-ACR) yielded a 237-bp amplicon and exhibited 90% (26/29) positive results with respect to A. caviae. None of the primer sets exhibited cross-reactivity with 12 non–A. caviae isolates and 52 other non-Aeromonas bacteria. The detection limit using the ACF2-ACR and ACF3-ACR primer sets in pure culture was 1.6?×?10³ CFU/mL, or 6 CFU per reaction, whereas that of the ACF1-ACR primer set was 1.6?×?10⁴ CFU/mL, or 60 CFU per reaction. In the case of spiked Nile Tilapia Oreochromis niloticus, the sensitivity of all primer sets without enrichment was 1.8?×?10⁴ CFU/g, or 30 CFU per reaction. Primer set ACF3-ACR was the best for a PCR assay targeting the gyrB gene, and the PCR technique developed was rapid, specific, and sensitive for the identification of A. caviae.

Received December 2, 2014; accepted April 9, 2015     

Oral Vaccination of Channel Catfish against Enteric Septicemia of Catfish Using a Live Attenuated Edwardsiella ictaluri Isolate

Enteric septicemia of catfish (ESC), caused by Edwardsiella ictaluri, is the most problematic bacterial disease affecting catfish aquaculture in the southeastern United States. Efforts to develop an effective ESC vaccine have had limited industrial success. In commercial settings, ESC vaccines are typically administered by immersion when fry are transferred from the hatchery to rearing ponds. While this approach is a practical method of mass delivery, this strategy administers vaccines to very young fish, which lack a fully developed immune system. To circumvent this limitation, an oral vaccination strategy was evaluated as a means of immunizing catfish at the fingerling stage of production, when fish possess a more complete immune arsenal. A virulent E. ictaluri isolate (S97-773) was attenuated by successive passage on media containing increasing concentrations of rifamycin. In laboratory trials, cultured vaccine was diluted and mixed with feed (100 mL diluted vaccine/454 g feed). This mixture was then fed to Channel Catfish Ictalurus punctatus fingerlings. Two separate dilutions of cultured vaccine (1:10 and 1:100) were used to create the vaccine–feed mixture, equating to estimated doses of 5 × 10⁷ and 5 × 10⁶ CFU/g of feed, respectively. After 30 d, catfish were exposed by immersion (1 × 10⁶ CFU/mL) to the virulent parental strain of E. ictaluri. The target dose (1:100 dilution, ～5 × 10⁶ CFU/g of feed) offered exceptional protection (relative percent survival = 82.6–100%). In addition, negligible deaths occurred in fish vaccinated at 10 times the target dose (1:10 dilution, ～5 × 10⁷ CFU/g of feed). In pond trials, antibody production increased 18-fold in orally vaccinated fish. When compared with nonvaccinated controls, vaccination significantly improved survival, feed fed, feed conversion, biomass produced, and total harvest. This research demonstrates Channel Catfish can be successfully immunized in a commercial setting against E. ictaluri with a single dose of an orally delivered, live attenuated, E. ictaluri vaccine.

Received July 31, 2014; accepted March 2, 2015     

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.     

Prevalence of and risk factors for Campylobacter colonisation in broiler flocks at the end of the rearing period in France

1. A study was conducted to estimate the prevalence and quantification by species of Campylobacter infection in broiler flocks at the end of the rearing period and to identify associated risk factors.

2. A questionnaire about the rearing period was completed and caecal samples were collected from 121 broiler flocks in Brittany, France, during 2008.

3. Campylobacter was isolated in 87 out of 121 flocks – a prevalence of 71.9% (95% CI, 63.7–80.1%), including 40.5% of Campylobacter jejuni and 29.8% of Campylobacter coli.

4. The average concentration, in positive flocks, was 7.96 log₁₀ cfu/g and ranged from 3.15 to 10.32 log₁₀ cfu/g.

5. The average concentration by species was: 7.57 log₁₀ cfu/g for C. jejuni and 8.44 log₁₀ cfu/g for C. coli.

6. There was a seasonal effect, with increased risk of Campylobacter colonisation in June, July and August (odds ratio (OR) = 9.59, 95% CI 1.15–79.75).

7. The other factors, associated with lower risk of Campylobacter colonisation, were the acidification of drinking water (OR = 0.33, 95% CI 0.13–0.86), antibiotic treatment at the beginning of the rearing period (OR = 0.20, 95% CI 0.07–0.55) and rodent control around the house (OR = 0.18, 95% CI 0.03–0.95).

8. The results show that hygiene practices and biosecurity measures could lead to a reduction in Campylobacter colonisation.     

An energy response curve for a commercial white leghorn laying‐strain cross

1. Individually‐caged White Leghorn hens, 235‐d‐old, were given daily metabolisable energy (ME) intakes ranging from 707 to 1321 kj for 8 two‐week periods. Energy was the first limiting nutrient, in those cases where the differences in egg output between treatments were sufficiently large.

2. Body weight, egg number and egg weight all responded (P < 0–001) to energy intake, and as judged by regression analyses, these responses had stabilised by the fifth period.

3. For a near‐maximum egg output of 48 g/bird d, the difference between the ME requirement of the average bird and of the flock, estimated from linear and curvilinear models respectively, was 20–5%.

4. The ME requirement (Y, kj/bird d) of the average bird for egg production (E, g/bird d) and maintenance of metabolic body size (kg W°. ⁷⁵ ) corrected to an ambient temperature of 22 °C is given by the equation, Y = 440W ^0.75 + 8.96 E     

Bioprocess enhancement of feather degradation using alkaliphilic microbial mixture

1. The main aim of this work is to develop a robust method to generate a microbial mixture which can successfully degrade poultry feathers to overcome environmental problems.

2. Four different alkaliphilic microbes were isolated and shown to degrade poultry feathers.

3. Two of the isolates were phylogenetically identified as Lysinibacillus and the others were identified as Nocardiopsis and Micrococcus.

4. The best microbial co-culture for white and black feather degradation was optimised for pH, temperature and relative population of the isolates to achieve almost 96% of degradation compared with a maximum of 31% when applying each isolate individually.

5. The maximum activity of keratinase was estimated to be 1.5 U/ml after 3 d for white feathers and 0.6 U/ml after 4 d for black feathers in a basal medium containing feather as the main carbon source. Additionally, non-denaturing polyacrylamide gel electrophoresis showed 4 and 3 protease activity bands for white and black feather, respectively.

6. This study provides a robust method to develop potential new mixtures of microorganisms that are able to degrade both white and black feathers by applying a Central Composite Design.     

Changes in structure of the limiting membrane and in oxygen permeability of the chicken egg integument during incubation

1. Oxygen permeabilities (K_O2 ) of the shell and shell membranes of fertile and infertile chicken eggs were measured at 37.5 °C and a relative humidity of 0.60 throughout 14 d incubation, with turning. The K_O2 of the shell and membranes of infertile eggs was around 1.0 × 10^?7 cm³ O₂STP sec^?1 cm^?2 Torr^?1 (1 Torr = 133.322 Pa) throughout incubation. With fertile eggs, from which there was a linear loss of water during incubation, the K_O2 of the shell and shell membranes was about 1.0 × 10^?7 cm³ O₂STP sec^?1 cm^?2 Torr^?1 for the first four days of incubation. Thereafter the majority of shells and membranes had a K_o2 of about 1.0 × 10^?6 cm³ O₂STP sec^?1 cm^?2 Torr^?1.

2. A diminution of the Na⁺ and K⁺ content of the shell membranes of fertile eggs was not associated with changes in the dimensions of the glyco‐protein mantle on the cores of the individual fibres of the membranes. There was, however, a progressive deterioration in the limiting membrane of fertile but not of infertile eggs.

3. It was concluded that changes in the O₂ resistance of the integument of fertile eggs were not a product of change in either of the shell membranes but of damage caused to the limiting membrane by the chorioallantois.     

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.     

Potential of a commercially available water acidification product for reducing Campylobacter in broilers prior to slaughter

1. This study investigated the potential of a commercially available acidified water treatment (PWT) for reducing the number of Campylobacter in vitro and other bacteria in the gut of live broilers.

2. In vitro tests indicated that PWT was highly effective for reducing Campylobacter jejuni and Campylobacter coli at the recommended concentration in water, reducing populations by greater than 7 log₁₀ CFU/ml after 24 h exposure. The decrease in the number of Salmonella serovar Enteritidis and Escherichia coli was not significant.

3. Addition of PWT to the broiler drinking water for the first 7 d, 2 d before and 2 d after each feed change and at feed withdrawal prior to slaughter or only after feed withdrawal had no effect on the number of Campylobacter in caecal samples on farm before thinning and depopulation compared to untreated controls.

4. Although PWT was effective for reducing Campylobacter in water, the results suggest that it does not reduce the number of Campylobacter in the caeca of broilers prior to slaughter under the conditions used in the study.     

Effect of TEX-OE® treatment on the development of heat shock proteins in commercial broiler chicks and the impact on performance indicators in the grow-out period

1. Heat shock proteins (HSPs) are highly conserved proteins, shown to protect organisms against physical and physiological stress.

2. TEX-OE^® is a patented total extract of the fruit of Opuntia ficus indica, which has been demonstrated to accelerate the development of HSPs in several animal species.

3. One-day-old commercial broiler chicks were treated with TEX-OE^®; HSP was measured by enzyme-linked immunosorbent assay (ELISA), and a large commercial field trial investigated key performance indicators (KPIs) in treated versus untreated controls chicks.

4. TEX-OE^® significantly increased HSP concentrations in treated chicks versus controls. Final cumulative mortality, liveweight and percentage factory-rejects were better than in controls.

5. The accelerated HSP response may enable chicks to cope with early stressors, which is reflected in improved KPIs.     

Effect of fermented moist feed on performance,gut bacteria and gut histo-morphology in broilers

1. Fermented feed has been shown to be beneficial in pig nutrition as a tool to reduce gut microbial disorders. Experiments with fermented feed in poultry are scarce, probably because of the belief that wet feed is less suitable for this species and causes wet litter.

2. A total of 280 one-d-old Ross 308 chickens were used in a completely randomised design with two dietary treatments (7 replicates of 20 birds/treatment); air-dry feed versus the same feed in moist form (water:feed ratio of 1.3:1, on a weight basis), inoculated with Lactobacillus plantarum NCIMB 40087 (9 log₁₀ CFU/kg feed) and batch-fermented for 48 h at 26°C. The birds were given starter (d 0–13), grower (d 4–26) and finisher (d 27–39) diets ad libitum. At the end of the grower and finisher period, two birds per pen were removed to sample intestinal contents for cultivating bacteria and intestinal tissue to determine villus height and crypt depth.

3. Fermented moist feed (FMF) batches showed good characteristics, with a pH between 3.9 and 4.4 and DL-lactic acid between 137 and 286 mmol/l. Daily feed intake and gain were reduced considerably in the FMF group in the starter (–40 and –44%, respectively) and grower (–23 and –16%) period, though in the finisher period these birds performed better, with an improved feed utilisation. Concomitant with the latter, villus height at the mid-jejunum and mid-ileum on d 39 was higher (+22.6% and +16.0%). Significantly more Lactobacilli and less coliforms were found in the foregut and less Streptococci in ileum and caeca of birds given FMF.

4. This trial showed that FMF was detrimental for early bird growth but affected beneficially feed efficiency, the composition of the gut bacteria and villus height in the small intestine in the finisher period in broilers.