Control of eight predominant Eimeria spp. involved in economic coccidiosis of broiler chicken by a chemically characterized essential oil.

AIM: To control eight most predominant Eimeria spp. involved in the economic disease of coccidiosis in broiler chicken, by a chemically characterized essential oil of eucalyptus and peppermint. METHODS AND RESULTS: The experimental design consisted of 160 day-old-broiler chicks, divided into four equal groups (G1 , G2 , G3 and G4 ), with 40 birds per group. Each group was divided into four equal subgroups. Birds in G1 were deprived of essential oil treatment and of Eimeria challenge. Birds in G2 were unchallenged, and administered the essential oil in drinking water at 0.69 ml kg(-1) body weight. Birds in G3 were untreated with essential oil, and each of its four subgroups was challenged at a different age (14, 21, 28 and 35 days). Birds in G4 were treated with essential oil, and challenged in the same manner as for G3 . Equal number of birds from all subgroups (n = 10) were sacrificed at the sixth day after the time allocated for each challenge. The 6 day incubation period post challenge resulted in respective mean per cent weight increase in G2 and G1 birds equivalent to 57.8 and 53.1% (P < 0.05). In addition, the essential oil improved the per cent weight increase in challenged birds (54.6%) compared to the challenged-untreated birds (18.6%) (P < 0.05). The mean feed conversion, mortality, intestinal lesion scores and oocyst counts were significantly reduced in the challenged-treated birds compared to the challenged-untreated birds (P < 0.05). CONCLUSIONS: The results support the hypothesis of using the essential oils of eucalyptus and peppermint to control the most prevalent Eimeria spp. involved in coccidiosis of broiler chicken, helping in improvement of their production, alleviation of lesions and reduction in intestinal oocyst counts. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information about the possibility of using this blend of essential oil as a coccidiostat for the protection of broiler chickens against the prevalent eight Eimeria spp. of coccidiosis.

Modulation by essential oil of vaccine response and production improvement in chicken challenged with velogenic Newcastle disease virus.

AIM: To evaluate the impact of Eucalyptus and peppermint essential oils on immune modulation and production of broiler chicken challenged with a molecularly characterized velogenic NewCastle disease virus (vNDV). METHODS AND RESULTS: The experimental design included five treatments with three replicate pens/treatment comprised of 12-day-old broilers chicks/replicate. The five treatments included a positive challenge control (non-NDV vaccinated/nonessential oil treated/challenged) (NNEOC), a negative challenge control (NDV vaccinated/essential oil treated/unchallenged) (VEOU), a non-NDV vaccinated/essential oil treated/challenged (NEOC), a NDV vaccinated/nonessential oil treated/challenged (VNEOC) and a NDV vaccinated/essential oil treated/challenged (VEOC). The lowest mean survival rate (0·0%) and lowest production performance were obtained by the positive challenge control, while the best mean survival (93·3%) and average body weight (2649 g) were obtained by the negative challenge controls (P < 0·05). Among the three others challenged treatments, the best mean survival (79·2%), highest mean body weight at 42 days of age (2445 g), the lowest feed conversion ratio (1·60) and the highest serum conversion immunopotentiation at 35 days of age determined by ELISA and hemagglutination titres were obtained by the VEOC birds compared with respective means obtained by birds of the NEOC and VNEOC treatments (P < 0·05). CONCLUSIONS: The results supported the possibility of using the essential oils of Eucalyptus and Peppermint in broilers to immunopotentiate the response to vaccination against velogenic NDV, helping in significant improvement of survival and production. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information about the potential use of essential oils of eucalyptus and peppermint that can be exploited as commercial immunopotentiators for the protection of NDV-vaccinated broiler chickens against economic velogenic NDV.

Anti-inflammatory bioactivities in plant extracts.

The medical ethnobotanical knowledge propagated over generations in the coastal regions of the Eastern Mediterranean, including Lebanon, is one that has built on several ancient cultures and civilizations of these regions. Recent interest in medical ethnobotany and the use of medicinal herbs in treating or preventing ailments has rejuvenated interest in folk medicine practices, especially those transcendent across generations. According to Eastern Mediterranean folk medicine practices, herbal remedies that treat many inflammation-related ailments were typically based on plant bioactive water extracts or decoctions. Studies have shown that active anti-inflammatory ingredients in water extracts include many natural chemicals such as phenols, alkaloids, glycosides, and carbohydrates. The intent of this manuscript is twofold: first, to review the literature that describes anti-inflammatory bioactivities in plant extracts of different plant genera; and second, to evaluate indigenous folk remedies used by folk doctors to treat inflammatory ailments in this region of the world. For this aim, the reported literature of five plant genera assumed to possess anti-inflammatory bioactivities and typically prescribed by folk doctors to treat inflammation-related ailments is reviewed.

Microbiological and chemical properties of litter from different chicken types and production systems.

Chicken litter is produced in large quantities from all types of poultry raising activities. It is primarily used for land application, thus it is essential to analyze its properties before it is released to the environment. The objective of this study is to compare the microbiological and chemical properties of litter generated from layer and broiler chickens reared under intensive and free-range production systems. The microbiological analysis consisted of the enumeration of total bacteria, total coliforms, Staphylococcus species, Salmonella species and Clostridium perfringens. Chicken litter from layers reared under intensive and free range systems showed lower mean total bacterial count than the litter collected from chicken broilers reared under either of the two systems (P=0.0291). The litter from intensive layers had the lowest mean total coliform counts (P=0.0222) while the lowest Staphylococcus species count was observed in the litter from free-range layers (P=0.0077). The C. perfringens count was the lowest in chicken litter from intensively raised broilers and layers (P=0.0001). The chemical properties of litter from the different chicken types and production systems were compared based on determination of pH, electrical conductivity, carbon, nitrogen, phosphorus, potassium, cadmium and zinc. Litter from free-range broilers showed the highest pH value (P=0.0005); however, the electrical conductivity was higher in the litter from both intensive and free-range layers compared to the litter from both broiler production systems (P=0.0117). Chicken litter from intensive systems had higher nitrogen content than litter from free-range systems (P=0.0000). The total phosphorus was the lowest in free-range broiler litter (P=0.0001), while the total potassium was the lowest in litter from intensively managed broilers (P=0.0000). Zinc appeared higher in litter from layers compared to that from broilers (P=0.0101). The cadmium content was higher in the litter from free-range broilers and layers compared to that in the litter from intensively managed systems (P=0.0439). Staphylococcus species in the litter as well as cadmium concentrations seem to be the most critical parameters presenting risks on the environment and on human health. Based on the lowest coliform counts (an indication of water pollution), the high nutrient levels and the low cadmium values, litter from intensively managed layers appears as the most suitable for application on agricultural soils.

Comparison of Mycoplasma gallisepticum subunit and whole organism vaccines containing different adjuvants by western immunoblotting.

Chickens were vaccinated with subunit (adhesin protein) or whole organisms of Mycoplasma gallisepticum (MG) adjuvanted with multilamellar positively charged liposomes or oil-emulsion. Sera were collected before and following the first (13 weeks of age) and second (17 weeks of age) vaccination. The chicken sera were used in western immunoblotting against whole MG polypeptides. Vaccination with the subunit (MG-adhesin) bacterin containing positively charged liposomes resulted in antibody response specific to adhesin band (75 kD) at 3 weeks post the first and second vaccination; however, crossreactions of the same antibodies occurred to MG proteins of 85 kD (3 weeks after the first vaccination) and 56 kD (3 weeks after the second vaccination). Vaccination with whole MG proteins containing positively charged liposomes resulted in significant immunopotentiation of antibodies against low molecular weight polypeptides of MG (less than 48.0 kD). The addition of Salmonella typhimurium cell wall proteins mitogens (STP) to the different bacterins suppressed the antibody responses to some MG polypeptides.

Preliminary data on efficacy of Mycoplasma gallisepticum vaccines containing different adjuvants in laying hens.

Chickens were vaccinated subcutaneously twice, at 13 and 17 weeks of age. The vaccines used were the whole organisms of Mycoplasma gallisepticum (MG) adjuvanted with multilamellar positively charged (MPC) liposomes or oil-emulsion. Other chickens received the same bacterins but supplemented with Salmonella typhimurium cell wall protein mitogen (STP) (50 micrograms/dose). At 21 weeks of age, each bird was challenged in the right and left caudal thoracic air sacs. The challenge dose/chicken was 1.3 x 10(5) CFU of MG (R-strain). A significant immunoglobulin (Ig) response specific to MG was observed in sera of chickens collected 3 weeks after the first and second vaccination with MG adjuvanted with MPC liposomes or oil-emulsion. The same two treatments had highly significant MG-titers in eggs collected during the first and second month post challenge. Both groups had highly significant protection (P less than 0.05) against MG transmission in eggs layed during the first month post challenge. Vaccination with MG organisms adjuvanted to MPC liposomes or oil-emulsion resulted in higher egg production, during the first month following challenge, in comparison to the unvaccinated-challenged birds; the same two groups had higher egg production in the second month following challenge compared to unvaccinated-challenged birds, but not significantly different (P greater than 0.05). The addition of STP to bacterins containing MG organisms adjuvanted to MPC liposomes or oil-emulsion, resulted in a significant reduction (P less than 0.05) of the Ig-specific to MG in sera and in a significant drop in egg production (P less than 0.05) during the first month following challenge.

Identification of the antigenic components of paramyxovirus-3, paramyxovirus-6 and Newcastle disease virus in turkeys.

The aim of this study was to use the enzyme-linked immunosorbent assay (ELISA) and the Western immunoblotting as possible tools to differentiate infections in turkeys by different paramyxoviruses. Pooled hyperimmune sera of turkeys infected with either paramyxovirus-3 (PMV-3), paramyxovirus-6 (PMV-6), or Newcastle disease virus (NDV) were assayed for antibodies specific to the three viruses by the ELISA and Western immunoblotting. ELISA results showed cross reactions of turkey antibodies between PMV-3 and PMV-6 antigens, while turkey antibodies to NDV did not cross-react with any of the other paramyxoviruses. The immunoblots of sera from birds infected with PMV-3 (Minnesota turkeys and Iowa chickens) reacted to low molecular weight polypeptides of PMV-3 of 29, 32, and 34 kDa, and to a high molecular weight band of 200 kDa. The same Minnesota turkey sera had a cross reaction to the 200 kDa polypeptide of PMV-6, while the Iowa chicken sera did not. Both sera had no apparent reaction to NDV proteins. Western immunoblotting showed that the turkey PMV-3 sera had a specific reaction to a 220 kDa polypeptide present in PMV-3, but not in PMV-6, while the turkey PMV-6 sera had a specific reaction to a 130 kDa polypeptide present in PMV-6, but not in PMV-3. Immunoblots of pooled sera from turkeys infected with PMV-6 (Minnesota source) reacted to the 200 kDa protein present in both PMV-3 and PMV-6; however, no reaction occurred between this sera and NDV proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of cell culture propagated and in vivo propagated hemorrhagic enteritis vaccines in turkeys.

The immune response of turkeys to a liquid, was compared with a previously frozen, cell culture propagated hemorrhagic enteritis (HE) vaccine. The liquid cell culture propagated HE vaccine was able to induce 100% seroconversion in turkeys 4 weeks after being vaccinated at 3.5 weeks of age; however, the previously frozen cell culture propagated HE vaccine induced 80% seroconversion 4 weeks post vaccination (P < 0.05). The average seroconversion in turkey flocks administered the liquid cell culture propagated HE was 97% in comparison with 98.5% in flocks given the splenic vaccine (P > 0.05). The complete absence of HE antigens in spleens of birds 5 days after being challenged with the virulent HE virus (40,000 TCID50 per bird) at an age of 9.5 weeks, was used as a model for successful protection against HE disease. The HE antigens were absent from spleens of all challenged birds that were previously vaccinated by the liquid cell culture propagated HE vaccine or splenic vaccine.

Identification of the antigenic components of the virulent Mycoplasma gallisepticum (R) in chickens: their role in differentiation from the vaccine strain (F).

The antibody response to different proteins of Mycoplasma gallisepticum (MG) was studied in chickens experimentally infected with virulent MG R strain. The chickens were challenged at 8 weeks of age by the intranasal route. Each cockerel received 1.3 X 10(6) colony-forming units (CFU). MG strains (R and F) were banded by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). The banding pattern was distinctively different between the two strains in the range of 92.5 to 200 kilodaltons (kD). Chicken sera collected at different times following challenge were analyzed by Western blot to determine the patterns of antibodies raised to specific MG proteins (R versus F strains). Early in infection (2 weeks postchallenge), antibodies to 60-kD and 75-kD polypeptides of MG R strain were produced. Subsequently (greater than or equal to 4 weeks postchallenge), antibodies recognized a larger number of MG antigens in both strains. The immunoblot patterns remained the same in the period 8-11 weeks postinfection in each of the two strains; however, the patterns were different when the two strains were compared. The early response recognized the 75-kD protein in the R strain while it recognized the 80-kD protein in the F strain. The late response recognized the 130-kD protein and the protein slightly heavier than 200 kD in the R strain. These two bands did not appear in the immunoblot performed against the F strain of MG. Electroeluted protein of MG R strain, namely adhesin (75 kD), showed a hemagglutination activity (HA) on chicken red blood cells. With the appearance of antibodies specific to the 60-kD and 75-kD polypeptides, there was a significant rise in hemagglutination-inhibition geometric mean titer of chicken sera.

Evaluation of an enrofloxacin-treatment program against Mycoplasma gallisepticum infection in broilers.

Three chicken broiler breeder flocks, 7 months of age, were confirmed to have Mycoplasma gallisepticum (MG) infection, based on culture of tracheal swabs. A total of fifty-five 7-day-old embryos from the three MG-positive flocks had an average 27.4% prevalence of MG-infection in their vitelline membrane. Sixty randomly selected MG isolates (30 from individual tracheas of breeders and another 30 from individual vitelline membrane of embryos) were highly sensitive in vitro to enrofloxacin (100%). Three broiler flocks (averaging 15,000 birds per flock) from the same three MG-infected chicken boiler breeders were divided into halves. The first halves were subjected to an enrofloxacin-treatment program and the other halves were controls. Sera collected at different ages of the broiler flocks were tested by the enzyme-linked immunosorbent assay for antibodies to MG. The absence of MG titers at 45 days of age in birds subjected to the enrofloxacin-preventive program was compared to an average prevalence of 15.9% in the controls (p < 0.05). The lack of MG titers in 45-day-old birds subjected to the enrofloxacin-treatment program was associated with lower better feed-conversion ratios (p < 0.05).

Isolation of salmonella and some other potential pathogens from two chicken breeding farms in Saudi Arabia.

Different salmonella serotypes were recovered from various sources in meat (A) and layer (B) poultry-breeder farms of Saudi Arabia. On farm A, salmonella were recovered from intestines of poultry breeders (7.41%), inshelled chicken embryos (2.87%), day-old chicks (19.23%), and mice (21.43%). Salmonellae in shell contents and on shell surfaces of hatching eggs contaminated the same percentage of eggs (1.24%), and litter and feed contaminations were 4.0% and 1.67%, respectively. On farm B, salmonella were recovered from intestines of poultry breeders (27.59%), mice intestines (15.0%), shells of hatching eggs (2.48%), hatching egg contents (0.35%), litter (33.33%), and feed (5.0%). No salmonella were recovered from intestines of inshelled chick embryos or day-old chicks on farm B. The total number of salmonella serotypes isolated from both farms was 14, and the serotypes common to both farms were Salmonella concord and S. livingstone. Salmonella contamination of shells of hatching eggs, pooled from both farms, was higher for dirty floor eggs than for clean eggs collected from floors and nests. The percentages of hatching eggs penetrated by bacteria other than salmonella on farms A and B, respectively, were: dirty floor eggs (25.53%, 22.83%), clean floor eggs (2.67%, 2.06%) and clean nest eggs (0.00%, 1.08%). The bacterial flora isolated from egg contents of unincubated hatching eggs (in decreasing order of frequency) were Staphylococcus, coliforms, Streptococcus, Bacillus, Salmonella, and Proteus.

Production of H2S by Escherichia coli isolated from poultry: an unusual character useful for epidemiology of colisepticemia.

Eleven isolates of H2S-producing Escherichia coli were recovered from necropsy materials of chickens with symptoms and lesions of colisepticemia on Saudi Arabian broiler farms. Results of 19 out of 20 biochemical reactions studied were typical for E. coli. Hydrogen sulfide production by the E. coli isolates was used as an epidemiological marker to pinpoint a breeding farm as the probable source of these strains, which were then transferred to progeny farms, where colisepticemia occurred. This finding was confirmed by the presence of the same antigenic structure (O78:H-) and by the same drug-resistance pattern (a multiple resistance to streptomycin, sulfathiazole, and tetracycline) in the isolates.

Distribution of paratyphoids on Saudi Arabian poultry farms and pathogenicity studies of predominant serotypes.

A total of 412 feed samples and 632 litter samples from 15 poultry farms (2 breeding farms and 13 rearing farms) were examined for salmonella. Twelve of these farms had salmonella in litter, five farms had salmonella in the feed and four had salmonella in both feed and litter. Seventeen feed samples (4.13%) and 121 litter samples (19.15%) were contaminated with salmonella. Sixteen salmonella serotypes were encountered, of which six were found in both feed and litter. Salmonella concord and S. livingstone were present in the litter of one breeding farm and its progeny farms. The five most frequently isolated salmonella serotypes in feed and litter were S. concord (17.39%), S. coeln (15.94%), S. livingstone (15.22%), S. manhattan (11.59%), and S. paratyphi B var. java (8.69%). The pathogenicities of those serotypes were determined by calculating their median lethal doses (LD50) 24 and 48 hr postinjection of 1,050 one-day-old broiler chicks via the navel into the yolk sac. The composite 48-hr LD50s (viable cells) were: S. concord, less than 8.8 X 10(3); S. livingstone, 1.1 X 10(5); S. manhattan, 3.5 X 10(5); S. coeln, 1.25 X 10(7); and S. paratyphi B var. java, 1.73 X 10(7).

New biotin-conjugated antisera for quantitation of Mycoplasma gallisepticum-specific immunoglobulin A in chicken.

The biotinylation of goat anti-alpha-chains of chicken immunoglobulin A (IgA), suitable for use in an avidin-biotin enzyme-linked immunosorbent assay, is described. The optimum conditions for the use of the developed conjugate in determining local and systemic IgA specific to Mycoplasma gallisepticum in chickens were established.

Characteristics of Actinomyces pyogenes involved in lameness of male turkeys in north-central United States.

A new condition of clinical lameness in 20 male turkey flocks of North-Central United States, associated with isolation of gram-positive rod bacteria from lesions of osteomyelitis, is characterized. The characterization confirmed the randomly selected isolates as Actinomyces pyogenes based on macroscopic and microscopic observations and 17 biochemical tests. The disease was reproduced within 3 weeks in all male turkeys, following an intravenous challenge at 15 weeks of age. The agar gel precipitin test and immunoblotting confirmed the antigenic similarity of the isolates recovered from the osteomyelitis lesions of lame birds.

An enzyme-linked immunosorbent assay for detection of Bordetella avium infection in turkey flocks: sensitivity, specificity, and reproducibility.

An enzyme-linked immunosorbent assay (ELISA) for detection of Bordetella avium infection in turkey poults was developed. One-week-old poults challenged intratracheally with 10(12) colony-forming units of B. avium had detectable titers (greater than or equal to 11), with an average of 13.6% positive samples when the birds were 6 to 11 weeks old. The method was sensitive enough to detect maternal antibodies to B. avium in poults up to 3 weeks of age. The same poults challenged at 1 week of age had 100% tracheal infection up to 3 weeks of age, which dropped to 0% by 6 weeks. The method resulted in no false-positive samples (titer = 0) from birds not infected with B. avium and tested weekly between 4 and 11 weeks of age. Antibodies in turkey flocks infected with Newcastle disease virus, hemorrhagic enteritis virus, and Mycoplasma meleagridis, and birds infected with Escherichia coli had no apparent cross-reactivity to the B. avium antigens used in the ELISA. The percentages of B. avium-positive serum samples collected from different turkey flocks did not significantly differ (P greater than 0.05) when samples were tested by the developed ELISA at different times, an indication of the reproducibility of the method.

Protection and immunity in commercial chicken layers administered Mycoplasma gallisepticum liposomal bacterins.

Six liposomal Mycoplasma gallisepticum (MG) bacterins, differing in charge and size, and two oil-emulsion vaccines (sonicated and non-sonicated) were given to white leghorns in two doses, at 13 weeks and again 1 month later. At 21 weeks of age, all chickens were challenged with a viable 20-hour culture of MG cells (17,800 colony-forming units) intratracheally and with nonviable MG organisms (0.09 mg protein) injected subcutaneously in the wattle center. The three chicken groups that had the lowest tracheal MG-infection rates postchallenge were those given adjuvants of small multilamellar positively charged liposomes (16.67%), large multilamellar negatively charged liposomes (16.67%), and non-sonicated oil-emulsion bacterin (37.5%). These three groups also had significant levels of antibody in sera 4 weeks after the second dose of vaccine. The group given the small multilamellar positively charged liposome also showed significant delayed-type hypersensitivity (wattle swelling) (P less than or equal to 0.05). The group given the large multilamellar negatively charged liposomes had the highest local antibody response (P less than or equal to 0.01) and was the only group that had no microscopic lesions in the trachea.

Serological response in broiler chicks to different commercial Newcastle disease and infectious bronchitis vaccines.

Broiler chicks were administered vaccines against Newcastle disease and infectious bronchitis (both Arkansas and Massachusetts strains) at 2 weeks of age as either primary or secondary vaccinations. The vaccine was administered as a spray at 2 weeks of age to chicks that had received Newcastle disease vaccine alone, bronchitis vaccine alone, both vaccines in combination, or no vaccine at day 1 in the hatchery. The Newcastle disease hemagglutination-inhibition response was significantly lower in chicks receiving Newcastle disease vaccine as a secondary vaccine at 2 weeks than in those receiving the vaccine as a primary vaccination at that age. In contrast, the bronchitis hemagglutination-inhibition response was significantly higher in chicks receiving bronchitis vaccine as a secondary vaccination at 2 weeks than in those receiving the vaccine as a primary vaccination at that age.

Immune response to Newcastle disease virus in broilers: a useful model for the assessment of detoxification of ervil seeds.

Ervil (Vicia ervilia) seeds are produced in the Mediterranean region and used as a source of protein for cattle and poultry. The methods used to assess the toxic effects of legume seeds in the feed of poultry include the observation of one or more parameters, including abnormal signs, weight gain, feed consumption, gall bladder weight, pancreas weight, pancreas proteolytic and amylase activity, haemolysis of red blood cells, liver weight, liver glutathione level, liver and plasma lipid levels, and plasma lipid peroxide levels. The authors describe the use of quantitative determination of the immune response to Newcastle disease virus (NDV) vaccine in broilers as a model that can be used to assess different detoxification treatments of ervil seeds. Broiler chicks fed differently-treated ervil, supplemented as 25% of the diet, at one to four weeks of age and vaccinated intraocularly with live NDV vaccine at eight days of age, showed different immune responses at three weeks post vaccination. Immunosuppression with regards to NDV was apparent in the group of birds raised on untreated ervil supplement, resulting in a mean immune response (titre) of 798.5. Five of the six different treatments of ervil seeds resulted in different degrees of rectification of the immunosuppression, with some broilers reaching a mean NDV immune titre of 2070.6, similar to that obtained in control broilers raised on a basal diet with no ervil seed supplement (mean NDV immune titre of 2333.8; P > 0.05). The five successful treatments of ervil (in increasing order of rectification of immunosuppression in broilers, with mean NDV titres in parentheses) were: ground soaked dried ervil (971.6), ground autoclaved dried ervil (1223.1), soaked autoclaved dried ervil (1273.1), soaked dried ervil (1340.0), and ground-soaked autoclaved dried ervil (2070.6).