Effect of dietary treatments on performance, oocysts shedding and lesion scores in broiler chickens experimentally challenged with Eimeria infection.

Although anticoccidials effectively control coccidiosis, a needed reduction in the reliance on antimicrobials in animal production leads to the exploration of alternative compounds. The present study aimed to test five different dietary treatments to counteract the negative impact of coccidiosis on broiler chickens' health and performance. 1-day-old male Ross 308 broilers (n = 960) were randomly assigned to one of eight treatments, with six cages per treatment (20 birds/cage). To the diet of the broiler chickens of treatments (Trt) 1-5, a synbiotic was added from d0-10. From d10-28, birds of Trt1 and Trt2 were fed synbiotics, whereas birds of Trt3 were fed diets with glutamine, and birds of Trt4 and Trt5 were fed diets with a combination of ß-glucans and betaine. From d28-35 onwards, birds of Trt1 were fed a diet with a synbiotic, whereas birds of Trt2-4 received diets with glutamine, and birds of Trt5 were fed a non-supplemented diet. Birds of the positive control group (PC; Trt6) were fed a standard diet supplemented with an anticoccidial (Decoquinate). The challenged negative control (NCchall; Trt7) and non-challenged negative control (NC) Trt8 were fed a standard diet without anticoccidial or other dietary treatment. At 7 days (d) of age, all birds were inoculated with 1 023, 115, and 512 sporulated oocysts of E. acervulina, E. maxima, and E. tenella, respectively, except for Trt8. Body weight gain (BWG), feed intake, and feed conversion ratio were assessed for each feeding phase (d0-10, d10-28 and d28-35) and overall experimental period (d0-35). Oocyst shedding, Eimeria lesion scores, cecal length, and relative weight were assessed at d13, d22, d28 and d35. Additionally, oocyst shedding was determined at d9 and d17. Litter quality was evaluated at d27 and d34, and footpad lesions at d34. During the starter (d0-10) and finisher (d28-35) periods, performance did not differ between the treatments. During the grower period (d10-28), Trt6 (PC) and Trt8 (NC) chickens had the highest BWG of all treatments (P < 0.001). Dietary treatment had no effect on litter quality and severity of footpad lesions. In the PC group (Trt6), low oocyst excretion and lesion scores were found. When comparing Trt1-5 with NCchall (Trt7), none of the treatments significantly reduced oocyst output or lesion scores. In conclusion, in this experiment, none of the dietary treatments performed similar or better compared to the PC group (Trt6) regarding performance or reducing Eimeria oocyst shedding or lesion scores.

Nutritional interventions to support broiler chickens during Eimeria infection.

Different combinations of gut health-promoting dietary interventions were tested to support broilers during different stages of Eimeria infection. One-day-old male Ross 308 broilers (n = 720) were randomly assigned to one of 6 dietary treatments, with 6 pens per treatment and 20 birds per pen, for 35 d. At 7 d of age (d7), all birds were inoculated with 1000, 100, and 500 sporulated oocysts of E. acervulina, E. maxima, and E. tenella, respectively. A 4-phase feeding schedule was provided. The dietary treatments (TRT) 1 to 4 included the basal diet supplemented with multispecies probiotics from d0 to 9 and coated butyrate and threonine from d28 to 35 but received four different combinations of prebiotics and phytochemicals from d9 to 18 and d18 to 28. The basal diet for the positive control (PC, TRT5) included diclazuril as a anticoccidial. The negative control (NC, TRT6) contained no anticoccidial. Performance was assessed for each feeding phase, and oocyst output, Eimeria lesion scores, cecal weight, litter quality, and footpad lesions were assessed at d14, d22, d28, and d35. Body weight gain (BWG) and feed intake (FI) were not affected by dietary treatment. PC broilers had the best feed conversion ratio (FCR) of all treatments from d0 to 35 (P < 0.001). None of the dietary treatments resulted in better litter quality or reduced footpad lesions compared to the PC. Moreover, the PC was most effective in reducing oocyst output and lesion scores compared to all other treatments. However, broilers that received the multispecies probiotics (d0 to 9), saponins (d9 to 18), saponins, artemisin, and curcumin (d18 to 28), and coated butyrate and threonine (d28 to 35) had the best FCR (P < 0.001) and lowest oocyst output and lesion scores compared to other dietary treatments. This study suggests that although the tested compounds did not perform as well as the anticoccidial, when applied in the proper feeding period, they may support bird resilience during coccidiosis infection.

Response to a Salmonella Enteritidis challenge in old laying hens with different vaccination histories.

Extending the laying period of laying hens is beneficial for economic and sustainability purposes. Because vaccines were designed with a shorter laying period envisaged, it is unclear whether current Salmonella vaccines can provide sufficient levels of protection against infection at an older age. The purpose of this experiment was to determine the efficacy of early rearing vaccination schemes against Salmonella challenge late in the laying period. There were four treatment groups: birds that had not been vaccinated (Group 1), birds vaccinated with live Salmonella Enteritidis (SE) (Group 2), with live and inactivated SE (Group 3), or with live SE and live Salmonella Typhimurium (Group 4). At the end of the laying period, the birds were transported from the laying farm to the research facility where they were orally challenged with 2.06 × 109 colony-forming units SE at around 82 wk of age. Hens were euthanized and bacteriology was performed on cecum, liver, spleen, and follicular fluid samples to determine SE colonization 7 and 14 d after challenge. Clinical and bacteriological findings of hens vaccinated with different vaccination schemes and the non-vaccinated control group were compared. No significant differences in SE colonization were found for vaccinated groups compared to the non-vaccinated control group. This may be a result of waning immunity due to the long time between vaccination and challenge. Also, as vaccination took place in the rearing period in the field, initial levels of immunity may not have been optimal due to shortcomings in the vaccination technique. Furthermore, the results of this study may have been affected by differences in age, breed, and origin between the groups. Therefore, controlled studies from early age onwards are necessary for more accurate comparisons between vaccines.

Enterococcus hirae-associated endocarditis outbreaks in broiler flocks: clinical and pathological characteristics and molecular epidemiology.

BACKGROUND: Enterococcus hirae-associated endocarditis, characterized by a peak in mortality during the second week of the grow-out, and occasionally lameness, was diagnosed at Dutch broiler farms. OBJECTIVES: Field cases were studied to increase knowledge on clinical and pathological characteristics, pathogenesis and epidemiology of these infections. ANIMALS AND METHODS: In total, 1266 birds of 25 flocks from 12 farms were examined. Post-mortem examinations, bacteriology, histopathology, PCR and DNA fingerprinting was carried out. Six flocks were followed longitudinally (n = 1017 birds). RESULTS: Average mortality was 4.1% for the entire grow-out, of which 36% was attributed to endocarditis. Fibrinous thromboendocarditis of the right atrioventricular (AV) valve was found in 24% of hearts, compared to 7% and 4% with lesions of left and both AV valves, respectively. Thrombotic lesions were found in 24% (n = 432) of lungs, but only in larger branches of the Arteria pulmonalis. Occasionally, thrombi were found in the Arteria ischiadica externa and in liver and brain vessels. Enterococcus was cultured from 54% (n = 176) of heart and in 75% (n = 28), 62% (n = 106) and 31% (n = 16) of liver, bone marrow and lung samples, respectively. Further identification, using the Rapid ID Strep 32 API system and a PCR targeting mur-2 and mur-2(ed) genes was carried out on a subset of Enterococcus positive isolates (n = 65): both techniques identified the isolates as Enterococcus hirae. Pulsed-field gel electrophoresis did not indicate evidence of clonality between farms and flocks. CONCLUSIONS: The relevance of these findings for pathogenesis and epidemiology of E. hirae infections is discussed. CLINICAL IMPORTANCE. This study may facilitate diagnosis of field cases and may contribute to the design of further research and development of control measures.

Efficacy of allicin from garlic against Ascaridia galli infection in chickens.

The use of garlic as a treatment against helminth infections is increasing in organic layer farms in several European countries. Its efficacy against these parasites, however, has not been demonstrated thus far. Therefore, a study was conducted to determine the efficacy of a commercially available garlic product consisting of a high concentration of allicin (i.e., the main active component of garlic) against experimentally induced Ascaridia galli infection in chickens. In total, 450 Lohmann LSL-Classic cockerels were used. Group 1, the uninfected, untreated group, consisted of 50 chickens. Groups 2 to 5, each consisting of approximately 100 chickens, were inoculated with 300 embryonated A. galli eggs/chicken at 6 wk of age. Group 2 was not treated, whereas groups 3 through 5 were given daily individual oral treatments from 13 wk of age onward. Group 3 received the recommended dose of allicin for 2 wk, whereas group 4 received a 10-fold dose of allicin. Group 5 was given 10 mg of flubendazole/kg of BW for 1 wk. Necropsy of 20 birds of all groups was performed weekly between 13 and 16 wk of age to determine adult worm loads. Group 1 remained free of A. galli. The experimental infection in the other groups resulted in a mean adult worm load of approximately 16 worms/bird. No significant differences were observed in worm counts of the allicin-treated groups (groups 3 and 4) compared with the infected, untreated group (group 2) at any week (P > 0.05). In contrast, no worms were found in chickens after flubendazole treatment (group 5). It was concluded that allicin does not represent an alternative to flubendazole for the treatment of A. galli infections in chickens.

Effect of Eimeria acervulina infection history on the immune response and transmission in broilers.

Heterogeneity in exposure to Eimeria spp. of chickens in a flock will result in differences between individual birds in oocyst output and acquired immunity, which subsequently affects transmission of the parasite in the population. The aim of this study was to quantify effects of previous infection of broilers with Eimeria acervulina on immune responses, oocyst output and transmission. A transmission experiment was carried out with pair-wise housed broilers, that differed in infection history. This "infection history" was achieved by establishment of a primary infection by inoculation of birds with 50,000 sporulated E. acervulina oocysts at day 6 of age ("primed"); the other birds did not receive a primary infection ("naïve"). The actual transmission experiment started at day 24 of age: one bird (I) was inoculated with 50,000 sporulated oocysts and was housed together with a non-inoculated contact bird (C). Oocyst excretion and parameters describing transmission, i.e. the number of infected C birds and time passed before start of excretion of C birds, were determined from day 28 to day 50 for six pairs of four different combinations of I and C birds (I-C): naïve-naïve, naïve-primed, primed-naïve and primed-primed. Immune parameters, CD4(+), CD8(+), αßTCR(+) and γδTCR(+) T cells and macrophages in duodenum, were determined in an additional 25 non-primed, non-inoculated control birds, and in the naïve-naïve and naïve-primed groups, each group consisting of 25 pairs. Although the numbers of CD4(+) T cells and γδTCR(+) T cells increased after primary infection, none of the immunological cell types provided an indication of differences in infectivity, susceptibility or transmission between birds. Oocyst output was significantly reduced in primed I and C birds. Transmission was reduced most in the primed-primed group, but nonetheless transmission occurred in all groups. This study also showed that acquired immunity significantly reduced oocyst output after inoculation and contact-infection, but not sufficiently to prevent transmission to contact-exposed birds.

Eimeria acervulina: the influence of inoculation dose on transmission between broiler chickens.

The course and clinical appearance of an Eimeria species infection in chicken flocks depend on the response of an individual bird to infection and on population-dynamics of the infection in the flock. Differences in ingested numbers of oocysts may affect oocyst load in the flock and the subsequent infectious dose for not yet infected birds. To study the link between numbers of oocysts excreted by infected birds and transmission of Eimeria acervulina, experiments were carried out with 42 pairs of broiler chickens using inoculation doses with 5, 50, 500 or 50,000 sporulated oocysts. In each pair one bird was inoculated and the other bird was contact-exposed. All contact birds became infected, which occurred on average within 34h after exposure to an inoculated bird. Although a higher inoculation dose resulted in higher oocyst excretion in inoculated and contact-infected birds, only small non-significant differences in transmission rates between groups were found.

Quantification of Eimeria acervulina in faeces of broilers: comparison of McMaster oocyst counts from 24h faecal collections and single droppings to real-time PCR from cloacal swabs.

Coccidiosis is an economically important disease in chickens, caused by infection with Eimeria species parasites. Diagnosis of coccidiosis is frequently based on oocyst enumeration in pooled faecal samples or litter. In studies on infection dynamics and for monitoring in the field, samples from individual chickens may be more appropriate as these support the determination of infection status of individual birds and more accurately reflect oocyst output at time of sampling. Faecal samples from individual birds can be collected, but the counting procedure limits the number of samples that can be processed and unequivocal microscopic differentiation between Eimeria species is very difficult. A test that overcomes these drawbacks would improve efficiency and quality of the diagnosis. The aim of this study was to compare two methods for Eimeria oocyst quantification in samples from individual birds. A real-time PCR that quantifies oocysts in cloacal swabs (qPCR) and oocyst counts in single droppings were compared to the standard procedure of oocyst counts in bulked 24h faeces. Faecal samples were collected daily from 30 broiler chickens, inoculated with different doses of Eimeria acervulina. The three techniques produced comparable oocyst counts for all inoculation doses. Single dropping counts are applicable for small sample sizes and when a single Eimeria species is used. For larger sample sizes qPCR is preferable as it can be carried out on samples that have been frozen for storage. Furthermore, qPCR can identify and quantify different Eimeria species, which makes it a valuable diagnostic tool for field or experimental work.

Heartworm in a mute swan (Cygnus olor).

A severely emaciated female adult mute swan (Cygnus olor) was euthanized after a period of intensive supportive care in a wild bird rehabilitation centre. Necropsy revealed severe myocardial haemorrhages in the right ventricular free wall associated with the presence of adult heartworm (Sarconema eurycerca). On histological examination, multifocal randomly scattered blood-filled cavities without endothelial or epithelial lining (migration tracts) and surrounded by necrotic debris mixed with fibrinoid deposits and a mixed inflammatory infiltrate were seen, as well as mild multifocal degeneration and necrosis of adjacent myocardial fibres. In some of the lesions, cut sections of adult female filarial nematodes revealed numerous microfilariae in their uteri. This report is the first published case of Sarconema eurycerca in a mute swan in The Netherlands.

Transmissibility of infectious bronchitis virus H120 vaccine strain among broilers under experimental conditions.

The aim of this study was to quantify transmission of infectious bronchitis virus (IBV) H120 vaccine strain among broilers, and to assess whether birds that have been exposed to vaccine strain-shedding birds were protected against clinical signs after infection with a virulent strain of the same serotype. A transmission experiment and a replicate were carried out, each with six groups of commercial broilers. At day of hatch (n = 30) or at 15 days of age (n = 20), half of each group was inoculated with either IBV H120 vaccine (H120 group), virulent IBV M41 (M41 group), or were mock-infected, thereby contact-exposing the other half of each group. Nasal discharge was recorded, and antibody response and virus shedding were measured. To measure clinical protection, four weeks after inoculation all birds, in all groups, were challenged with IBV M41. The reproduction ratio (R; the average number of contact infections caused by one infectious bird) was determined to quantify virus transmission. All contact-exposed birds, except for one in an H120 group, became infected with either IBV H120 or IBV M41. Almost all birds contact-infected with IBV H120 or IBV M41 were subsequently protected against clinical signs after challenge with IBV M41. The lower limits of the 95% confidence interval (CI) of the R of IBV H120 vaccine, and of IBV M41, were significantly <1. For both IBV H120 and IBV M41, the 95% CI was [2.1-infinity] following inoculation at day of hatch and [1.8-infinity] after inoculation at 15 days of age. This finding demonstrates that IBV H120 vaccine is able to spread extensively among broilers. This implies that this vaccine strain might be able to become endemically present in the poultry population. It also implies that, even if not all birds received vaccine during spray application, due to the ability of the vaccine to spread in the flock, they will most likely be protected against clinical signs after a subsequent field virus infection.

Outbreak of avian influenza H7N3 on a turkey farm in the Netherlands.

This case report describes the course of an outbreak of avian influenza on a Dutch turkey farm. When clinical signs were observed their cause remained unclear. However, serum samples taken for the monitoring campaign launched during the epidemic of highly pathogenic avian influenza in 2003, showed that all the remaining turkeys were seropositive against an H7 strain of avian influenza virus, and the virus was subsequently isolated from stored carcases. The results of a reverse-transcriptase pcr showed that a H7N3 strain was involved, and it was characterised as of low pathogenicity. However, its intravenous pathogenicity index was 2.4, characterising it as of high pathogenicity, suggesting that a mixture of strains of low and high pathogenicity may have been present in the isolate. The outbreak remained limited to three farms.

Isopathic and pluralist homeopathic treatment of commercial broilers with experimentally induced colibacillosis.

This study sought to determine the efficacy of isopathic and pluralist homeopathic treatment of colibacillosis in broiler chickens and thereby contribute to the evaluation of homeopathy in general. In each of two experiments three groups of broilers, infected intratracheally at 8 days of age with E. coli (O78:K80), were treated with different combinations of homeopathic remedies. Control groups and an infected, doxycyline-treated group were included. Experiments differed only in the dose of E. coli. Efficacy of treatment was evaluated based on the parameters mortality, body weight gain and colibacillosis lesions. In both experiments doxycyline prevented mortality and reduced E. coli lesions and stunting. None of the homeopathically treated groups differed significantly with respect to any of the parameters from the non-medicated, infected control group. It is concluded that the results of this study do not justify use of these homeopathic remedies for treatment of colibacillosis in broilers. Furthermore, no significant effects of this homeopathic treatment were established.