Lack of Protection following Re-Exposure of Chinook Salmon to Ceratonova shasta (Myxozoa)

The recent identification of multiple genotypes of the salmonid parasite Ceratonova shasta with different virulence levels in Chinook Salmon Oncorhynchus tshawytscha suggests that it is possible to immunize fish against subsequent infection and disease. We hypothesized that exposure of Chinook Salmon to the less-virulent parasite genotype (II) prior to the more virulent parasite genotype (I) would decrease disease and/or result in fewer mature parasites compared with fish only infected with the more virulent genotype. To test this hypothesis, fish were challenged in a combination of field and laboratory exposures, and we measured infection prevalence, percent morbidity, and mature parasite production. Neither mortality nor mature parasite production were reduced when fish were exposed to genotype II prior to genotype I compared with fish exposed only to genotype I, suggesting that protection against C. shasta using a less-virulent genotype of the parasite does not occur.

Received June 9, 2014; accepted September 19, 2014     

Effect of Nanophyetus salmincola and Bacterial Co-Infection on Mortality of Juvenile Chinook Salmon

The freshwater trematode Nanophyetus salmincola has been demonstrated to impair salmonid immune function and resistance to the marine pathogen Vibrio anguillarum, potentially resulting in ocean mortality. We examined whether infection by the parasite N. salmincola similarly increases mortality of juvenile Chinook Salmon Oncorhynchus tshawytscha when they are exposed to the freshwater pathogens Flavobacterium columnare or Aeromonas salmonicida, two bacteria that juvenile salmonids might encounter during their migration to the marine environment. We used a two-part experimental design where juvenile Chinook Salmon were first infected with N. salmincola through cohabitation with infected freshwater snails, Juga spp., and then challenged with either F. columnare or A. salmonicida. Cumulative percent mortality from F. columnare infection was higher in N. salmincola-parasitized fish than in nonparasitized fish. In contrast, cumulative percent mortality from A. salmonicida infection did not differ between N. salmincola-parasitized and nonparasitized groups. No mortalities were observed in the N. salmincola-parasitized-only and control groups from either challenge. Our study demonstrates that a relatively high mean intensity (>200 metacercariae per posterior kidney) of encysted N. salmincola metacercariae can alter the outcomes of bacterial infection in juvenile Chinook Salmon, which might have implications for disease in wild fish populations.

Received February 24, 2015; accepted September 7, 2015     

Infecting Pacific Herring with Ichthyophonus sp. in the Laboratory

The protistan parasite Ichthyophonus sp. occurs in coastal populations of Pacific Herring Clupea pallasii throughout the northeast Pacific region, but the route(s) by which these planktivorous fish become infected is unknown. Several methods for establishing Ichthyophonus infections in laboratory challenges were examined. Infections were most effectively established after intraperitoneal (IP) injections with suspended parasite isolates from culture or after repeated feedings with infected fish tissues. Among groups that were offered the infected tissues, infection prevalence was greater after multiple feedings (65%) than after a single feeding (5%). Additionally, among groups that were exposed to parasite suspensions prepared from culture isolates, infection prevalence was greater after exposure by IP injection (74%) than after exposure via gastric intubation (12%); the flushing of parasite suspensions over the gills did not lead to infections in any of the experimental fish. Although the consumption of infected fish tissues is unlikely to be the primary route of Ichthyophonus sp. transmission in wild populations of Pacific Herring, this route may contribute to abnormally high infection prevalence in areas where juveniles have access to infected offal.

Received June 25, 2015; accepted September 12, 2015     

Natural resistance of Sri Lankan village chicken to Salmonella gallinarum infection

1. An experiment was conducted to compare the natural resistance of an indigenous breed of local village chickens to Salmonella gallinarum with two commercial breeds: ISA Brown and ISA White layers under experimental conditions.

2. A total of 72 chickens from each of these breeds were randomly distributed to 4 pens to provide equal numbers of two replicate pens maintained as infected and control (uninfected). All chickens in infected groups were inoculated orally with 1 × 10⁸ CFU (1 ml dose) of a field isolate of S. gallinarum, at the age of 8 and 16 weeks given over 5 consecutive days. Growth performance, clinical signs, gross pathological lesions and antibody responses were measured.

3. A significantly higher mortality was observed in the brown layers compared with the white layers, and clinical signs and mortality were absent in village chickens. However, a large number of birds with gross lesions and high antibody titres were detected in village chickens, indicating that birds had the disease subclinically. Commercial breeds had a significantly higher body weight, feed intake and feed conversion efficiency.

4. There was a significantly lower proportion of positive reactors in village chickens in the whole-blood agglutination test (35%) compared to brown (100%) and white (90%) layers even after the second inoculation. Uninfected birds were negative in all groups. The indirect enzyme-linked immunosorbent assay confirmed these observations.

5. These results suggest that the indigenous breed had superior natural resistance to S. gallinarum than the commercial breeds.     

Antitrichomonal activity of Peganum harmala alkaloid extract against trichomoniasis in pigeon (Columba livia domestica)

1. This study was designed to evaluate the antitrichomonal effects of P. harmala alkaloid extract against T. gallinae, both in vitro and in vivo, as well as comparing it to that of metronidazole, conventional antitrichomonal medication and harmine and harmaline, the two alkaloids present in P. harmala.

2. T. gallinae were collected by the wet mount method from infected free-living pigeons. The in vitro assay was performed using multi-well plates containing test compounds in final concentrations of 5, 10, 15, 20, 30, 50 or 100 μg/ml. The in vivo assay was done on 60 experimentally infected pigeons dosed with metronidazole at 50 mg/kg body weight (BW) or alkaloids at 25 mg/kg BW.

3. The 24 h minimum inhibitory concentration (MIC) of alkaloid extract was 15 µg/ml while that of metronidazole was 50 µg/ml. Harmine and harmaline revealed 24 h MIC of 30 and 100 µg/ml, respectively. Treatment of infected pigeons with alkaloids led to a full recovery after 3 d but with metronidazole total eradication of trophozoites was not achieved.

4. In conclusion, data of the present study suggested P. harmala is a potent natural anti-trichomonal agent, effective against T. gallinae.     

Effect of butyrate,clopidol and their combination on the performance of broilers infected with Eimeria maxima

1. The effect of butyric acid glycerides (BAGs) with and without clopidol (CLP) on Eimeria maxima on growth and associated biochemical variables was investigated in broiler chickens.

2. One-day-old chicks were divided into 6 equal groups (Gps) of 30 chicks each; each group was subdivided into 6 equal subgroups. Gp 1 was not infected and not treated. Chicks in Gp 2 were not infected and fed on a ration mixed with 4 g BAGs/kg for 6 successive weeks. Chicks of the other groups were directly inoculated intra-crop with 1 × 10⁴ sporulated oocysts of E. maxima at 14 d of age. Gp 3 was infected and not treated. Chicks in the remaining three groups were given diets mixed with the tested drugs for 6 successive weeks. Gp 4 was fed on a diet mixed with CLP (125 g/kg). Gp 5 was given a diet mixed with BAGs (4 g/kg diet). Gp 6 was fed on a diet mixed with both BAGs (4 g/kg diet) and CLP (125 g/kg).

3. Birds in Gps 5 and 6 showed a reduction in the mean oocyst count, lesion scores and developmental stages in the lamina propria and improved growth and biochemical variables. BAG supplementation enhanced growth and production of healthy broilers.

4. It was concluded that BAGs were a useful supplement in broiler diets as an alternative to growth promoters and antimicrobial drugs.     

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.     

Pharmacokinetics of difloxacin in healthy and E. coli-infected broiler chickens

1. The pharmacokinetics of difloxacin were investigated in healthy and E. coli-infected broiler chickens following intravenous and oral administration of a single dose of 10 mg/kg bodyweight.

2. After intravenous injection of difloxacin, the serum concentration–time curves were best described by a two-compartment open model. The distribution and elimination half-lives (t_0.5α) and (t_0.5el), respectively, were 0.10 ± 0.016 h and 3.7 ± 0.08 h in healthy chickens compared with 0.05 ± 0.005 h and 6.42 ± 0.71 h in E. coli-infected birds. The volumes of distribution V_dss were 3.14 ± 0.11 and 9.25 ± 0.43 l/kg, with total body clearance (Cl_tot) of 0.65 ± 0.018 and 1.14 ± 0.1 ml/kg/h, respectively.

3. Following oral administration, difloxacin was absorbed with t_0.5(ab) of 0.57 ± 0.06 and 0.77 ± 0.04 h and was eliminated with t_0.5(el) of 4.7 ± 0.34 and 3.42 ± 0.19, respectively, in normal and infected chickens. The peak serum concentrations were 1.34 ± 0.09 and 1.05 ± 0.06 µg/ml and attained a T_max of 2.27 ± 0.07 and 2.43 ± 0.06 h, respectively. The systemic bioavailability of difloxacin following oral administration was 86.2% in healthy chickens and 90.6% in E. coli-infected birds. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of difloxacin against the field strain of E. coli O78 in vitro were 0.02 µg and 0.04 µg/ml, respectively.

4. These results show that administration of a therapeutic dose of difloxacin is effective in the treatment of E. coli infection in chickens. The serum concentration of the drug was much higher than the MIC of the E. coli O78 strain in both healthy and infected chickens.     

Electroencephalographic assessment of concussive non-penetrative captive bolt stunning of turkeys

1. The aim of this study was to evaluate the electroencephalographic (EEG) and behavioural responses of turkeys stunned with three different concussive non-penetrative captive-bolt guns prior to slaughter.

2. A total of 31 slaughter weight ex-breeding female turkeys (29 weeks of age; mean body weight 13.32 ± SD 0.65 kg) were stunned with the Cash Poultry Killer (CPK) (n = 10), Turkey Euthanasia Device (TED) (n = 10) and Zephyr EXL (n = 11).

3. Mean peak kinetic energy was highest for the CPK compared to the TED and Zephyr EXL (75.9 ± 4.5, 28.4 ± 0.4 and 24.4 ± 0.7 J, respectively).

4. A total of 29 (94%) of the turkeys were rendered unconscious following captive bolt stunning, with total power of the EEG (Ptot) significantly reduced from baseline values (reductions of 67% CPK, 84% TED and 76% Zephyr EXL, p < 0.01) and waveforms becoming isoelectric after periods of transitional EEG. However, two birds shot with the CPK and Zephyr EXL had periods of behavioural/reflexes (rhythmic respiration, nictitating membrane reflex, neck tension) and EEG activity (43–47 and 36–60 + s after the shot, respectively) indicating incomplete concussion and return of consciousness. In one bird, the shot was incorrectly positioned (Zephyr EXL), while the other appeared to be related to a defective cartridge (CPK).

5. In conclusion, all three captive bolt gun models were effective in producing unconsciousness in turkeys, provided they were positioned correctly and power loads performed according to their specifications.     

Absorption and deposition of xanthophylls in broilers challenged with three dosages of Eimeria acervulina oocysts

1. An experiment was designed to evaluate the effect of different doses of oocysts of Eimeria acervulina on intestinal absorption and skin deposition of xanthophylls (XAs) in broilers.

2. A total of 192 broiler chickens were randomly assigned to 4 groups: an uninfected control group and three groups inoculated with either 1 × 10², 1 × 10⁴ or 1 × 10⁵ sporulated oocysts of E. acervulina by gavaging at 21 d. There were 4 replicate pens (2 male and 2 female) per group.

3. Plasma xanthophyll (PX) and skin yellowness (SY) were measured in live birds weekly. At 42 d of age, SY was measured in the breast and abdomen after chilling and in the breast 24 h post-processing on refrigerated carcasses.

4. In general, in all challenged treatments, and for the duration of the study, the average PX decreased by 0.02 μg/ml (R² = 61.6%) for every 1000 inoculated oocysts, whereas PX increased by 1.26 μg/ml/d in uninfected birds.

5. The average SY in live birds from 21 to 42 d of age decreased by 0.019 b*/every 1000 oocysts administered, while SY of uninfected controls increased by 0.57 b*/d. It was also noted that in all treatments females had a greater SY (6.17 b*) than males for the duration of the study. The SY of the breast and abdomen was correlated (r = 0.76) in chilled carcasses. Breast SY in 24 h refrigerated carcasses was greater in the control group and for female birds.

6. Oocyst excretion was different between inoculated treatments only on 7 d post-inoculation (PI). Coccidia lesion scores in the duodenum averaged 1+ in infected birds and 2+ in birds given the highest oocyst dose.     

The use of n‐paraffin‐grown yeast as the main source of protein in diets for chicks

1. n‐Paraffin‐grown yeast and a mixture of soyabean meal and fish meal were compared in the net protein utilisation (NPU) test, and as protein supplements in diets for broilers up to 4 weeks of age.

2. The difference between the NPU values, 0.66 for yeast and 0.80 for the soyabean meal and fish meal mixture, could largely be attributed to the high nucleic acid content of the yeast.

3. Chicks given the diet containing yeast (190 g/kg) did not grow as rapidly as those given the soyabean meal and fish meal reference diet, and the reduced growth could only partly be explained by a marginal deficiency of methionine.

4. Food conversion efficiency with the yeast diet was improved by maize oil while responses to α‐tocopheryl acetate and sodium selenite were inconsistent.

5. Chicks grew well when yeast replaced fish meal in the mixture of soyabean meal and fish meal, and when fish meal (194 g/kg diet) was the sole protein supplement.     

Detection of Infectious Pancreatic Necrosis Virus from the Leeches Hemiclepsis marginata and Hirudo medicinalis

Leeches have been reported to harbor several important fish pathogens, including spring viremia of carp virus, infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV), and also may contain blood protozoa. In the present study, leeches were collected from water bodies located in Kurdistan province, Iran. The specimens were tested for IHNV, VHSV, and infectious pancreatic necrosis virus (IPNV) using the PCR method. The results showed that two different species of leeches, Hemiclepsis marginata and Hirudo medicinalis, were infected by IPNV among the seven species studied. The infected leeches were found in areas that were polluted with untreated sewage coming from upstream fish farms culturing Rainbow Trout Oncorhynchus mykiss. In addition, the fish at fish farms in the vicinity had been infected with IPNV 9 months previously. Our results showed that the virus causing infectious pancreatic necrosis is present in the leeches H. marginata and H. medicinalis, suggesting that leeches are a potential source of IPNV in fish farms.

Received October 14, 2015; accepted June 1, 2016 Published online September 29, 2016     

Prevalence and antimicrobial resistance of Arcobacter species isolated from poultry meat in Iran

1. The objective of this study was to determine the prevalence and antimicrobial resistance of Arcobacter spp. isolated from different species of retail poultry meat in Iran.

2. From August 2012 to April 2013, a total of 540 raw poultry meat samples from chicken (n = 100), turkey (n = 100), quail (n = 100), partridge (n = 80), duck (n = 50), ostrich (n = 60) and geese (n = 50) were purchased from randomly selected retail outlets in Shahrekord, Isfahan, Sari and Rasht, Iran.

3. Using culture techniques, 71 of 540 poultry meat samples (13.1%) were positive for Arcobacter spp. The highest prevalence of Arcobacter spp. was found in chicken meat (28.0%), followed by quail (12.0%), duck (11.4%), turkey (11.0%), geese (8.0%), partridge (7.5%) and ostrich (3.3%) meat. The number of A. butzleri isolated from poultry meat samples (90.1%) was significantly higher than A. cryaerophilus (7.1%) and A. skirrowii (2.8%). Significantly more poultry meat samples were found to contain Arcobacter spp. by the PCR assay than by the culture method.

4. Susceptibilities of Arcobacter isolates were determined for 14 antimicrobial drugs using the disk diffusion method. All of the 71 Arcobacter isolates tested were resistant to one or more antimicrobial agents. Resistance to cephalothin and vancomycin (95.8%) was the most common finding, followed by resistance to methicillin, azithromycin and ampicillin. All Arcobacter isolates were susceptible to gentamicin, streptomycin, tetracyclin and kanamycin.

5. The results of this study indicated the importance of poultry meat, especially chicken meat, as potential sources of Arcobacter spp. infection in people. Furthermore, the strains indicated resistance to a broad spectrum of antibiotics.     

The occurrence of giant fibres in different muscles of two chicken genotypes

1. The occurrence of Giant Fibres (GF) in three muscles (Pectoralis major (PM), Iliotibialis lateralis and Semimembranosus) with different types of energy metabolism was studied in slow- and fast-growing chicken strains.

2. A total of 20 one-day-old Leghorn chicks (slow-growing) and 20 broiler (Ross 508) chicks (fast-growing) were reared to 100 and 45 d, respectively.

3. A small percentage of GF was seen in pre rigor muscle samples even at 3 min post mortem in both genotypes and in all muscle types studied.

4. From 3 min to 24 h post mortem GF increased both in Leghorn and broiler chickens but to a different extent according to muscle type and genotype.

5. The highest GF 24 h post mortem value was found in the PM muscles belonging to the fast-growing broiler line.

6. It was concluded that every type of muscle can develop GF, but this phenomenon is more evident in the PM especially in animals selected for increased growth rate.     

Effect of gender on breast and thigh turkey meat quality

1. The influence of gender on chemical composition, physicochemical parameters, fatty acid profile, amino acid and mineral composition of turkey breast and thigh meat was studied in order to assess nutrient requirements.

2. Chemical composition showed that only intramuscular fat in breast meat was significantly affected by gender (p < 0.05). The results showed a higher percentage of intramuscular fat in male samples, almost double the amount found in females (0.73% vs. 0.38%).

3.For meat colour parameters, only a* showed different results between sexes, with male samples (breast: p < 0.01; thigh: p < 0.001) having the highest values.

4. Fatty acid profiles showed that medium chain unsaturated fatty acids were the most abundant. The significant differences (p < 0.05) found in both breast and thigh muscle could be linked to a difference in metabolism between males and females.

5.There were higher levels of C16:1n-7 in females (breast: p < 0.001; thigh: p < 0.01) compared with male muscle sample (5.05 vs. 2.67 g/100 g in breast and 4.95 vs. 3.27 g/100 g in thigh). Nutritional indices (n-6/n-3 and thrombogenic index) were more favourable in female samples demonstrating that female turkeys had better fatty acid profile than the others.

6. Turkey meat is an important source of dietary amino acids, and female samples had the highest contents both of essential and non-essential amino acids. Furthermore, gender had a numeric effect (p > 0.05) on amino acid composition.

7. Mineral composition showed that Na, Zn and Fe were the minerals most affected by turkey gender.     

Prevalence of gastrointestinal helminth infections in free-range laying hens under mountain farming production conditions

1. A cross-sectional study was conducted from September 2015 to July 2016 in South Tyrol, Northern Italy to examine the prevalence of gastrointestinal helminths in free-range laying hens under mountain farming production conditions.

2. A total of 280 laying hens from 14 free-range mountain farms (4 organic, 10 conventional) were randomly collected at the end of the laying period. Faecal samples were taken to analyse faecal egg counts (FEC) and faecal oocyst counts (FOC). The gastrointestinal tracts were removed post mortem and examined for the presence of helminths.

3. In faeces, FEC values averaged 258 eggs per g of faeces, which were dominated by Ascaridia galli and Heterakis gallinarum. Mean FOC was 80 oocysts/g. In the gastrointestinal tract, at least one nematode species was found in 99.3% of the examined hens. H. gallinarum was the most prevalent nematode (95.7%), followed by Capillaria spp. (66.8%) and A. galli (63.6%). Thirty per cent of the chickens were infected with cestodes (tapeworms). Correlation coefficients between worm counts of H. gallinarum, Capillaria spp. and A. galli ranged from 0.41 to 0.51.

5. The helminth prevalence did not differ between conventional and organic farms, whereas total worm burden was higher in organic compared with conventional farms (318.9 vs. 112.0). Prevalence and infection intensity did not differ between farms that used anthelmintic treatments and those that did not.

6. In conclusion, free-range laying hens under the studied mountain farming conditions are at high risk of nematode infection, especially in organic systems. The vast majority of hens are subclinical infected with at least one helminth species.     

Persistence of salmonella enteritidis in poultry units and poultry food

1. Studies on the survival of Salmonella enteritidis in poultry units and food were carried out over a two‐year period.

2. The organism persisted for at least one year in an empty trial house at the laboratory in which naturally‐infected broiler breeder birds had previously been housed. A similar survival period was seen in a building which had housed an infected layer breeder flock, although infection was not detected in a subsequent pullet flock.

3. Salmonella enteritidis was also frequently found surviving outside poultry houses in small pockets of litter and fan dust which had been left after cleansing and disinfection of the site. On some poultry units S. enteritidis was also found in wild bird droppings.

4. Salmonella contamination appeared to persist preferentially in association with dust particles swept from the floor and in food troughs and S. enteritidis survived at least 26 months in artificially contaminated poultry food.     

Effects of zinc–methionine on growth performance,intestinal flora and immune function in pigeon squabs

1. Different concentrations of zinc–methionine (Zn-Met) were given to pigeon squabs, and the resulting effects on growth, immune functions and intestinal microflora were investigated from hatching to 28 d of age. A total of 180 artificially hatched pigeon squabs were randomly allotted to each of three treatments with three replicates of 20 squabs. The three treatments given were either one ml (2 mg/ml) Zn-Met, one ml (10 mg/ml) Zn-Met or one ml 0.9% NaCl solution.

2. The results showed that Zn-Met improved the growth performance of squabs. The average daily and average weekly weight gain was significantly greater in squabs treated with Zn-Met than in the control group.

3. The group given 2 and 10 mg supplemental Zn-Met had heavier thymus, spleen and bursa of Fabricius than the control group at d 28.

4. Maternal antibody titres against Newcastle disease haemagglutination inhibition and alpha-naphthyl acetate esterase were significantly higher in squabs treated with supplemental 2 and 10 mg Zn-Met compared to the control group at d 14 and d 28.

5. Additionally, the squabs given supplemental 2 mg Zn-Met exhibited significantly higher Bacillaceae, Lactobacillus, Enterococcus and Bifidobacterium populations at d 14 and d 28, but lower Escherichia coli populations at d 28 compared to the control group. On the contrary, Lactobacillus, Enterococcus and Bifidobacterium populations were significantly decreased with 10 mg Zn-Met at d 28.

6. This study indicates that supplementation with Zn-Met has a positive effect on growth performance, immune function and regulation of intestinal flora in pigeons. An inclusion level of 2 mg seems to be better than 10 mg Zn-Met per day per bird.