Occurrence of infection with Toxoplasma gondii and factors associated with transmission in broiler chickens and laying hens in different raising systems

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. The aim of the present study was to determine the occurrence and identify the risk factors associated with transmission of T. gondii to chickens raised in different systems (free-ranged and confined) to produce eggs or meat. The 810 animals were allocated in two experimental groups according to the production system purpose: 460 broiler chickens (Group 1) and 350 layer chickens (Group 2). In order to analyze the possible factors involved in T. gondii infection in the chickens, an epidemiological questionnaire was developed for all properties.The serological detection of anti-Toxoplasma gondii antibodies was performed by Indirect Immunofluorescence (IFAT) and by Enzime Linked Imunossorbent Assay (ELISA). Since the agreement index (kappa) between these two serological techniques was considered high, 21.2% of the 810 animals were considered reactive. In Group 1, 12.2% (56/460) were positive, while in the Group 2 the positivity rate was 33.1% (116/350). The production system may be influencing the seropositivity of the animals in both groups. However, only in Group 2 it was possible to notice a statistically significant relationship between the breeding system and the frequency of positive sera. This result indicates that, at least for laying hens, the production system is directly involved in T. gondii infection. The contact with cats in Group 1 did not influence the distribution of seroreactive animals, but in Group 2 a significant relationship was observed. The occurrence of anti-T. gondii antibodies was high in both groups (broiler and posture chickens). Free-ranged chickens raised for egg production proved to be the most exposed group to the T. gondii infection. This can be related to the fact that these animals stay for longer periods in the farms, in direct contact with possibly contaminated soil by the presence of domestic cats.

Effects of starter diet supplementation with arginine on broiler production performance and on small intestine morphometry

The effects of starter diet (days 1 to 21) supplemented with arginine (Arg) on the production performance and duodenum and jejunum mucosa morphometry of broilers were studied. Male Cobb broiler chickens (990) were randomly assigned to one of five treatments in a complete random design. Measurements of 33 chicks per treatment were made in six repetitions. The treatments consisted of a basal diet with 1.390% digestible Arg (no supplementation) and four dietary levels (1.490%, 1.590%, 1.690%, and 1.790%), providing a relationship with lysine of 1.103; 1.183; 1.262; 1.341 and 1.421%, respectively. From the age of 22 days on, all birds received conventional grower diet. The data were submitted to regression analysis by polynomial decomposition of the degrees of freedom in relation to the levels of Arg. The Arg supplementation increased (P<0.05) the live weight and the feed conversion ratio without increasing the feed intake of the birds. However, no effect was observed (P>0.05) in the growth phase (days 22 to 42) in the absence of the Arg supplementation. The supplementation of Arg over of NRC recommendation during the starter phase may be necessary for the expression of the maximal weight gain potential in birds. No effect (P<0.05) of Arg dietary supplementation was observed either on small intestine weight and length at any age. However, the duodenum villus:crypt ratio increased and the crypt depth decreased in the first week in response to increasing dietary Arg. It is concluded that broiler Arg dietary supplementation in the starter diet improved production performance and small intestine morphometry, especially in the first week.

Nested-PCR for the detection of Mycoplasma hyopneumoniae in bronchial alveolar swabs, frozen tissues and formalin-fixed paraffin-embedded swine lung samples: comparative evaluation with immunohistochemical findings and histological features

The diagnosis of Mycoplasma hyopneumoniae infection is often performed through histopathology, immunohistochemistry (IHC) and polymerase chain reaction (PCR) or a combination of these techniques. PCR can be performed on samples using several conservation methods, including swabs, frozen tissue or formalin-fixed and paraffin-embedded (FFPE) tissue. However, the formalin fixation process often inhibits DNA amplification. To evaluate whether M. hyopneumoniae DNA could be recovered from FFPE tissues, 15 lungs with cranioventral consolidation lesions were collected in a slaughterhouse from swine bred in herds with respiratory disease. Bronchial swabs and fresh lung tissue were collected, and a fragment of the corresponding lung section was placed in neutral buffered formalin for 48 hours. A PCR assay was performed to compare FFPE tissue samples with samples that were only refrigerated (bronchial swabs) or frozen (tissue pieces). M. hyopneumoniae was detected by PCR in all 15 samples of the swab and frozen tissue, while it was detected in only 11 of the 15 FFPE samples. Histological features of M. hyopneumoniae infection were presented in 11 cases and 7 of these samples stained positive in IHC. Concordance between the histological features and detection results was observed in 13 of the FFPE tissue samples. PCR was the most sensitive technique. Comparison of different sample conservation methods indicated that it is possible to detect M. hyopneumoniae from FFPE tissue. It is important to conduct further research using archived material because the efficiency of PCR could be compromised under these conditions.

Effects of fumonisin B1 on selected biological responses and performance of broiler chickens

The objective of this study was to determine the effects of three doses of fumonisin B1 (0, 100, and 200mg/kg of feed) on biological variables (relative weight of liver [RWL], total plasma protein [TPP], albumin [Alb], calcium [Ca], phosphorus [P], uric acid [UA], alanine aminotransferase [ALT], aspartate aminotransferase [AST], gamma glutamyltransferase [GGT], alkaline phosphatase [AP], total cholesterol [Chol], triglycerides [Tri], sphinganine-to-sphingosine ratio [SA:SO], and C-reactive protein [CRP]), morphological evaluation of the small intestine (villus height [VH], crypt depth [CD], and villus-to-crypt ratio [V:C]), histological evaluation, and on performance (body weight [BW], feed intake [FI], and feed conversion rate [FCR]) of broiler chickens. Significant effects of FB were observed on BW and FI (reduced), on RWL, TPP, Ca, ALT, AST, GGT, Chol, and Tri (increased) at both 14 and 28 days evaluations. In addition, significant increase was observed on FCR, Alb, P, SA:SO, and CRP and significant reduction in UA, VH, and V:C only at the 28 days evaluation. Significant histological lesions were observed on liver and kidney of FB inoculated broilers at 14 and 28 days. Those results show that FB has a significant effect on biological and histological variables and on performance of broiler chickens.

Induction of immune response in broiler chickens immunized with recombinant FliC and challenged by Salmonella Typhimurium

The study examined (1) the immune response in broiler chickens after oral immunization with recombinant flagellin (rFliC) from Salmonella Typhimurium conjugated with sodium alginate microparticles, and the immune response enhancement in association with recombinant cholera toxin B subunit protein (rCTB) and pool of Lactobacillus spp. (PL). The immune responses were evaluated by dosage of IgY serum and IgA from intestinal fluid and immunostaining of CD8+ T lymphocytes in the cecum. The immunized animals were challenged with Salmonella Typhimurium (ST) 21 days after treatment. In all immunized groups, a significant increase (p<0.05) was observed in IgA levels (μg/mL), especially three weeks after immunization. The serum IgY levels (μg/mL) were little affected by the treatments and differed significantly among groups only in the second post-immunization week (p<0.05). After the challenge, the number of CD8+ T cells differed significantly between the treatments and negative control. Retrieval of Salmonella Typhimurium was not detected at 48 hours after the challenge in T2 (rFliC+rCTb), T3 (rFliC+PL) and T4 (rFliC+rCTB PL). The rFliC administered orally with or without rCTB and Lactobacillus spp. produces significant induction of humoral immune response, and the immunized chickens were more effective in eliminating Salmonella after challenge.

Fowl adenovirus Group I as a causal agent of inclusion body hepatitis/hydropericardium syndrome (IBH/HPS) outbreak in brazilian broiler flocks

Commercial broiler flocks from a farm located in the State of São Paulo, Brazil, presented diarrhea, depression, increased mortality and poor weight gain. Upon post-mortem examination, classical signs of Inclusion Body Hepatitis/Hydropericardium Syndrome (IBH/HPS) were observed, including enlarged pale yellow-colored livers and straw-colored liquid in the pericardial sac. In addition, gross lesions were also observed in the kidneys, pancreas, thymus, intestines and gallbladder. Samples of these organs were analyzed by PCR for the detection of the hexon gene of the Fowl Adenovirus (FAdVs) Group I. The results were positive for both flocks (A and B) assayed by PCR. The macroscopic lesions associated with the detection of FAdV Group I by PCR in several of these affected organs allowed for the identification of IBH/HPS. In fact, this is the first report in Brazil of IBH/HPS in broilers, which identifies FAdVs group I as a causal agent of the disease. These findings may contribute to the worldwide epidemiology of the adenovirus-mediated hepatitis/hydropericardium syndrome.

Occurrence of Campylobacter jejuni and C. coli on broiler carcasses after chilling in southern Brazil

Campylobacter jejuniand C. colihave been associated with gastrointestinal disorders in human beings, due mainly to the consumption of chicken meat. Despite control measures for reducing contamination by these bacteria, the detection of Campylobacter in carcasses after chilling remains high.A total of 105 carcasses were assessed by the horizontal detection method in five federally inspected slaughterhouses in southern Brazil in 2012 and in the first three months of 2013. Campylobacterwas isolated in 37.1% of the carcasses, of which 97.5% contained C. jejuni and 2.5% were infected by C. coli. The rate of positive carcasses across the slaughterhouses ranged from 0 to 71.4%. Determining the occurrence of Campylobacteramong flocks is crucial for estimating the microbial load at specific points along the slaughtering process and for minimizing the risk of contamination of end products by Campylobacter.

Resistance to β-lactam and tetracycline in Campylobacter spp.isolated from broiler slaughterhouses in southern Brazil

Abstract The study was carried out to screen and analyze the genetic characteristics of antimicrobial resistance in Campylobacter spp. from poultry sources. A total of 141 strains of Campylobacter isolated from samples of broilers of slaughterhouses in southern Brazil was identified by phenotypic and genotypic methods. Campylobacter isolates were evaluated for its antimicrobial susceptibility and the presence of resistance genes. The strains were investigated for antimicrobial susceptibility against two agents (ampicillin and tetracycline) by disk diffusion method. PCR assay was used to confirm the specie and the presence of ampicillin (blaOXA-61), tetracycline tet(O), and the energy-dependent multi-drug efflux pump (cmeB) genes. Campylobacter jejuni was the most ubiquitous; its presence was determined in 140 samples out of 141 (99.3%), whereas Campylobacter coli was found only in one of the contaminated samples (0.70%). The results obtained showed 65% and 35.5% of Campylobacter isolates resistant to β-lactams and tetracyclines, respectively. The cmeB gene responsible for multidrug resistance was detected in 26 isolates out 141 strains (18.5%). Moreover, 36 out of 141 Campylobacter strains (25.6%) were found to be resistant to at least two different antimicrobia resistance markers (β-lactams and tetracyclines).

Effects of Cinnamon extract on biochemical enzymes, TNF-α and NF-κB gene expression levels in liver of broiler chickens inoculated with Escherichia coli

Abstract: Infection with Escherichia coli (E. coli) is a common disease in poultry industry. The use of antibiotics to treat diseases is facing serious criticism and concerns. The medicinal plants may be effective alternatives because of their multiplex activities. The aim of this study was to investigate the effects of cinnamon extract on the levels of liver enzymes, tumor necrosis factor-alpha (TNF-α) and nuclear factor-kappa B (NF-κB) gene expressions in liver of broiler chickens infected with E. coli. Ninety Ross-308 broilers were divided into healthy or E. coli-infected groups, receiving normal or cinnamon extract (in concentrations of 100 or 200mg/kg of food) supplemented diets. E. coli suspension (108cfu) was injected subcutaneously after 12 days cinnamon administration. Seventy-two hours after E. coli injection, the blood samples were taken for biochemical analysis of liver enzymes in serum (spectrophotometrically), and liver tissue samples were obtained for detection of gene expression of inflammatory markers TNF-α and NF-κB, using real-time PCR. Infection with E. coli significantly increased the levels of TNF-α and NF-κB gene expressions as well as some liver enzymes including creatine-kinase (CK), lactate-dehydrogenase (LDH), alanine-transferase (ALT) and aspartate-transferase (AST) as compared with control group (P<0.05). Pre-administration of cinnamon extract in broilers diet (in both concentrations) significantly reduced the tissue levels of TNF-α and NF-κB gene expressions and enzymes CK and ALT in serum of broiler chickens inoculated with E. coli in comparison with E. coli group (P<0.05 and P<0.01). The levels of LDH and AST were significantly decreased only by 200mg/kg cinnamon extract in infected broilers. The level of alkaline-phosphatase (ALP) was not affected in any groups. Pre-administration of cinnamon extract in diets of broiler chickens inoculated with E. coli could significantly reduce the gene expression levels of pro-inflammatory mediators and liver enzymes activities, thereby protecting the liver against this pathologic condition.

Physicochemical evaluation and fatty acids profile of broiler chicken fed broiler diets containing barley brewer (Hordeum vulgare)

The aim of this study was to analyze the chemical composition and the fatty acids (FA) profile of broiler chicken (breast and thigh/on-thigh) fed diets containing 10 and 20% of barley and a control diet. The FAs were evaluated by gas chromatography. There was not a significant difference (p > 0.05) in the breast chemical composition. In the thigh/on-thigh cut, the ration containing 20% of barley reduced the ashes (p < 0.05). Regarding the saturated FA composition, only the breast myristic acid treated with 10% of barley was reduced (p < 0.05). None of the diets influenced the polyunsaturated (PUFA) and monounsaturated (MUFA) FA's content (p > 0.05). As for the ω6/ω3 ratio, the highest one was observed in the 20% barley fed animals, in both cuts, being considered the worst quality. It can be concluded that the 20% barley ration was worse than the others and the 10% barley ration was better than the control ration, especially when considering the saturated FA quantity that was reduced.

Evaluation of temperatures in a refrigerated container for chilled and frozen food transport

The objective of this work was to monitor the operational conditions of the transport of chilled and frozen foods during delivery within cities and to evaluate the impact of the door openings on the alteration of the internal temperature of the refrigerated environment. Several temperature and pressure sensors were used in a refrigerated container with two compartments and they were installed in the refrigeration system unit and on the internal and external surfaces of the container. After the monitoring tests, it was verified that door openings during deliveries resulted in a disturbance that raised the internal temperature of the refrigerated container above values recommended for adequate conservation of the products transported. Moreover, increasing the number of door openings promoted a cumulative effect on the internal temperature, mainly in the chilled food compartment of the container. It was concluded that the refrigeration system unit presented serious limitations with regard to the maintenance of the container's internal temperature during the actual distribution routine, since it does not possess enough instantaneous capacity to restore the temperature set-point between deliveries.

Microstructural changes of frozen strawberries submitted to pre-treatments with additives and vacuum impregnation

Strawberries were submitted to freezing after pre-treatments with hydrocolloid and calcium salts (pectin and calcium chloride) at different concentrations, in the attempt to establish a correlation of the effects of these substances and their processing, on the physical and microstructural characteristics of fruits after thawing. Strawberry halves were submitted to impregnation with controlled vacuum pressure of 84.4, 50.5 and 16.6 kPa; comprising pectin at concentrations of 0, 1.5 and 3%; with the addition of calcium chloride at concentrations of 0, 3 and 6%; and glucose at 20%, for 4 hours. Measurements were made of the total soluble solid contents, cellular fluid loss, texture and viscosity of the solution, before and after the freezing/thawing. Images of the tissue cuts during the freezing, in function of time, were taken in an optic microscope coupled to a cold-stage and controlled temperature system, where the reduction of the cellular area was quantified using an image analyzing software. The pectin concentration had an influence on and demonstrated a potential for protection of the frozen tissue samples. The photomicrographs showed that the loss of cellular fluid occurs during the growth of ice formed in the intercellular spaces and it is retarded through treatments with high pectin concentrations.

Stability of frozen marolo pulp during storage

Marolo, also known as araticum or head-to-black, is a globular berry, a species native to the Brazilian savannah. The aim of this study was to evaluate the physical, chemical, and microbiological stability of frozen marolo pulp during 12 months of frozen storage. It was observed that the levels of ash (0.28-0.22%), protein (0.77-0.71%), lipids (1.75-1.73%), carbohydrates (12.1-10.15%), calorie (67.23-59.01 kcal), sucrose (2.50-1.29%), citric acid (435.63-197.5 µg.g-1), tartaric acid (4.38-1.88 µg.g-1) , acetic acid (470.38-279.25 µg.g-1), ascorbic acid (3.00-0.00 µg.g-1), total pectin (0.67-0.39%), pH (3.88-3.83), and b* chromaticity coordinates (24.85-20.53) decreased reduced during storage, whereas the levels of moisture (85.10-87.19%), color parameters (L* 58.89-62.62 and a* 5.37-7.86), reducing sugars (4.53-5.62%), total soluble sugars (7.1-7.36%), soluble solids (7.0-8.4 ºBrix), total acidity (0.9-1.0%), malic acid (514.13-781.25 µg.g-1), soluble pectin (0.16-0.24%), and antioxidant (6.85-37.35% of DPPH discoloration) increased over the one-year of storage period. According to the physical, chemical, and microbiological parameters assessed, the product can be stored for 12 months without loss of quality with addition of citric acid as a preservative.

Changes in the physicochemical and microbiological properties of frozen araça pulp during storage

Araça belongs to the Myrtaceae family and is popularly known as araçá-comum, araçá-azedo, or araçá-do-campo. Frozen fruit pulp is of great importance for the food industry, which can produce it at the time of harvest, store it, and use it according to the demand of the consumer market and/or as an ingredient in the formulation of products such as yogurt, candies, and ice creams among others. The aim of this study was to evaluate the physical, chemical, and microbiological stability of frozen araça pulp during 12 months of frozen storage. It was observed that the levels of moisture (90.55-88.75%), ash (0.34-0.26%) total soluble sugars (7.11-6.62%), sucrose (3.55-1.39%), soluble pectin (0.24-0.23%), total pectin (0.5-0.46%), pH (3.82-2.31%), organic acids (698.12-122.25 µg.g-1 citric acid), and phenolic compounds (6.22-0.00 mg GAE.100 g-1) decreased during storage, whereas the levels of protein (0.61-0.83%), lipids (0.14-0.38%), total carbohydrates (8.36-9.78%), calorific value (37.14-45.86 kcal.100 g-1), reducing sugars (3.51-5.21%), soluble solids (5.17-6.0%), total antioxidant capacity (6.89-35.13%), and color parameters (L*49.75-50.67; a*0.79-1.82 and b*22.5-25.19) increased over the one-year storage period. According to the chemical and microbiological parameters assessed, the product can be stored for 12 months without loss of quality with addition of citric acid as a preservative.

Correlation, by multivariate statistical analysis, between the scavenging capacity against reactive oxygen species and the bioactive compounds from frozen fruit pulps

The contents of total phenolic compounds (TPC), total flavonoids (TF), and ascorbic acid (AA) of 18 frozen fruit pulps and their scavenging capacities against peroxyl radical (ROO•), hydrogen peroxide (H2O2), and hydroxyl radical (•OH) were determined. Principal Component Analysis (PCA) showed that TPC (total phenolic compounds) and AA (ascorbic acid) presented positive correlation with the scavenging capacity against ROO•, and TF (total flavonoids) showed positive correlation with the scavenging capacity against •OH and ROO• However, the scavenging capacity against H2O2 presented low correlation with TF (total flavonoids), TPC (total phenolic compounds), and AA (ascorbic acid). The Hierarchical Cluster Analysis (HCA) allowed the classification of the fruit pulps into three groups: one group was formed by the açai pulp with high TF, total flavonoids, content (134.02 mg CE/100 g pulp) and the highest scavenging capacity against ROO•, •OH and H2O2; the second group was formed by the acerola pulp with high TPC, total phenolic compounds, (658.40 mg GAE/100 g pulp) and AA , ascorbic acid, (506.27 mg/100 g pulp) contents; and the third group was formed by pineapple, cacao, caja, cashew-apple, coconut, cupuaçu, guava, orange, lemon, mango, passion fruit, watermelon, pitanga, tamarind, tangerine, and umbu pulps, which could not be separated considering only the contents of bioactive compounds and the scavenging properties.