Abstract in English:ABSTRACT HMOX1 is an important gene in biosynthesis of the eggshell pigment of blue eggs. Previous studies found that HMOX1 is highly expressed in the shell gland of hens laying blue eggs (BlueH) compared with hens laying brown eggs (BrownH); however, the reasons for the differential expression are unclear. In this study five single nucleotide polymorphism (SNP) in HMOX1 were genotyped in 111 BlueH and 115 BrownH. The association of haplotypes of these SNP with the blue egg phenotype was tested. Haplotype-specific expression of HMOX1 was detected in the shell gland. The interaction of sequence variants and transcription factors was analyzed using electrophoretic mobility shift assay (EMSA). A TG haplotype covering upstream 1.4 kb region of HMOX1 was significantly associated with blue eggs (p<0.05). Furthermore, the birds (n=12) with the haplotype expressed 3.8 fold more transcripts than those (n=12) without the haplotype (p<0.05). After re-sequencing a 2.2 kb region harboring the TG haplotype, a total of 26 SNP were found, of which a SNP was predicted to create a binding site of Nrf2, a transcription factor initiating HMOX1 expression. However, subsequent EMSA failed to confirm the Nrf2-DNA interaction. Taken together, the data suggested that the TG haplotype is not directly involved in regulation of HMOX1 expression; a regulatory mutation located near the haplotype and linked with the haplotype may exist and be responsible for the differential expression ofHMOX1.
Abstract in English:ABSTRACT Avian metapneumovirus (aMPV) is a negative-sense single-stranded RNA enveloped virus of the Metapneumovirus genus belonging to theParamyxoviridae family. This virus may cause significant economic losses to the poultry industry, despite vaccination, which is the main tool for controlling and preventing aMPV. The aim of this study was to evaluate the antiviral activity of extracts of four different native plants of the Brazilian Cerrado against aMPV. The antiviral activity against aMPV was determined by titration. This technique measures the ability of plant extract dilutions (25 to 2.5 µg mL-1) to inhibit the cytopathic effect (CPE) of the virus, expressed as inhibition percentage (IP). The maximum nontoxic concentration (MNTC) of the extracts used in antiviral assay was 25 µg mL-1for Aspidosperma tomentosum and Gaylussacia brasiliensis, and 2.5 µg mL-1 for Arrabidaea chica and Virola sebifera. Twelve different extracts derived from four plant species collected from the Brazilian Cerrado were screened for antiviral activity against aMPV. G. brasiliensis, A. chica, and V. sebifera extracts presented inhibition rates of 99% in the early viral replication stages, suggesting that these extracts act during the adsorption phase. On the other hand, A. tomentosum inhibited 99% virus replication after the virus entered the cell. The biomonitored fractioning of extracts active against aMPV may be a tool to identify the active compounds of plant extracts and to determine their precise mode of action.
Abstract in English:ABSTRACT Salmonellosis is an infection caused by specific or non specific serotypes of theSalmonella genus, responsible for losses in the poultry industry. Fowl typhoid, caused by S. Gallinarum (SG) is important because it causes elevated mortality in adult birds, leading to economic losses in the poultry industry. This study aimed at quantifying the number of viable SG cells in the liver, spleen, lung, cecum, and reproductive tract (ovary and testicles) of experimentally inoculated Japanese quails (Coturnix coturnix), as well as SG shedding in their feces. One hundred and two Japanese quails, with four months of age at the beginning of the experiment, were used. The birds were inoculated with three bacterial cultures containing different concentrations (6x104 CFU/0.1mL, 2x105 CFU/0.4mL, or 5x106 CFU/0.2mL) of SG resistant to nalidixic acid. On days 1, 4, 7, and 14 after the inoculation (dpi) individual cloacal swabs were collected from six birds per group, which were subsequently sacrificed for organ sampling. The swab samples were streaked directly on plates containing brilliant green agar and nalidixic acid (VBNal). Samples that were negative after 24h, were streaked again. The collected organs were individually macerated and transferred to buffered peptone water at 0.1%. The solutions were immediately diluted serially for CFU counting in VBNal. SG was successfully recovered from one quail, which was inoculated with 2x105 CFU/0.4mL, and from five quails of the group inoculated with 5x106 CFU/0.2mL inoculum. All of the analyzed cloacal swab samples were negative. Therefore, this study demonstrated it was difficult to isolate SG from the analyzed organs and that it was not possible to recover thepathogen in the cloacal swabs collected from inoculated quails. These results may be explained by the absence of flagella in SG, inducing weak intestinal immune response in the beginning of the infection and preventing its isolation in cloacal swab samples. The low positivity rate of the analyzed organs may be due to the immune status of the euthanized birds, since the SG dissemination in the animal organism occurs mostly close to death, which was observed in the birds found dead during the experiment.
Abstract in English:ABSTRACT Intramuscular fat (IMF) content in chickens significantly contributes to meat quality. The main objective of this study was to assess the expression of calcineurin (CaN) and Ca2+/calmodulin-dependent protein kinase (CaMK) in lipogenesis in chicken muscle. Chickens were slaughtered and sampled at 4, 8, and 16 weeks of age. IMF content and the expression of CaN subunits and CaMK isoforms were measured in the thigh muscle tissue. The results showed that the IMF contents were greater at 16 weeks compared with those at 4 and 8 weeks (p<0.05). Transcription of fatty acid synthase (FAS) and fatty acid translocase CD36 (FAT/CD36) mRNA significantly increased with age, from four to 16 weeks (p<0.05). The mRNA levels of CaN B and CaMK IV were significantly lower at 16 weeks than at four weeks (p<0.05), but CaMK II mRNA levels were significantly higher than at four weeks (p<0.05). In order to evaluate the role of CaMK and CaN in adipogenesis, SV cells were incubated in standard adipogenic medium for 24 h and treated with specific inhibitor of CaMK and CaN. The expressions of CCAAT/enhancer binding protein b (C/EBPb, sterol regulatory element-binding protein 1 (SREBP1), and peroxisome proliferation-activated receptor g (PPARγ) were dramatically enhanced by the CsA, CaN inhibitor (p<0.05). KN93, CaMK II inhibitor, dramatically repressed the expression of those lipogenic gene (p<0.05). These results indicated that CaN and CaMK had different effects on adipogenesis in the muscle of chickens.
Abstract in English:ABSTRACT This study aimed at examining the effect of overfeeding on the activity of the mTOR pathway in the liver and muscle tissues of Gang geese. Eighty healthy male Gang geese were reared under the same feeding conditions, and were divided at 14 weeks of age into a control group and an overfed group. All birds were slaughtered after three weeks of over feeding. Gene expression and protein content of several genes involved in the mTOR pathway were evaluated. The results showed that the gene expression of mTOR, raptor, and rictor, and the protein contents of mTOR and PI3K were higher in liver, breast muscle, and leg muscle of the overfed group than that of control group. However, the S6K expression level was clearly lower in the liver of the overfed group than that of control group, and there was no evident difference in both breast muscle and leg muscle between the control group and the overfed group. These results suggest that overfeeding induces the activity of raptor, rictor, and mTOR, and that mTOR signaling pathway was closely linked with PI3K pathway in the evaluated geese.
Abstract in English:ABSTRACT The pre-slaughter period is considered critical in broiler production. Several factors contribute to increase the birds' stress, such as handling, harvesting, and transportation, negatively affecting their welfare. This study aimed at evaluating the addition of lemon grass (Cymbopogon citratusStapf) to the drinking water of broilers during the pre-slaughter period on their behavior, blood cortisol, and surface temperature. The study was carried out at the experimental farm of the Federal University of Grande Dourados (UFGD), Dourados, MS, Brazil. In total, 2594 broilers were distributed according to a completely randomized experimental design, in a 3x2x2 factorial arrangement, with four replicates per treatment. Treatments consisted of three different lemon grass levels (Cymbopogon citratus Stapf) used in the form of an infusion (0, 0.1, and 5 g per L of water), sex (male or female), and genetic strain (Ross(r) 308 or Cobb(r) 500). The infusion was offered when birds were 42 days old. On that day, blood was collected for blood cortisol level determination, broiler surface temperature was recorded, and an ethogram was applied to register broiler behavior. Blood cortisol level and broiler surface temperature were not affected by treatments (p>0.05). The behavior of beak opening was different between the genetic strains (p<0.05), being more frequent in Ross(r) 308 broilers. Lemon grass water content did not affect broilers' surface temperature when consumed during the pre-slaughter period.
Abstract in English:ABSTRACT This study was conducted to evaluate the nutritional values of fractions of damaged corn. One hundred and eighty 22-d-old Cobb 500 male broilers were distributed in batteries according to a completely randomized design with six treatments of six replicates each. The treatments consisted of diets containing five corn fractions, classified as sound, fermented, insect-damaged, mold-damaged, or reference corn. The test diets consisted of 60% of reference diet + 40% of each corn fraction. Only the reference corn fraction included all the fractions at different proportions (0.8% fermented, 0.05% insect-damaged, 3.3% mold-damaged, and 95.85% sound grains). The method of total excreta collection was used to determine AMEn values and metabolizability coefficients of dry matter (MDM), crude protein (MCP), ether extract (MEE), and gross energy (MGE) of the reference corn and its fractions. The density values of the corn fractions were used to calculate the correlations among the evaluated parameters. The evaluated corn fractions presented different compositions values. The insect-damaged and mold-damaged grains presented higher CP level, lower density, and MDM and MCP coefficients compared with the other fractions. However, calculated AMEn values were not significantly different (p>0.05) among corn fractions. A low correlation between density and AMEn content (r<0.5), and a high correlation between density and MCP (r>0.8) were calculated. Although the evaluated corn fractions presented different nutritional values, there were no marked differences in their utilization by broilers.
Abstract in English:ABSTRACT An experiment was conducted to evaluate the performance and nutrient metabolizability of broilers fed diets containing fumonisin B1 (FB1) and an esterified glucomannan (EGM). In total, 420 male broilers were distributed according to a 3 x 2 + 1 factorial arrangement, corresponding to three FB1 exposure times (seven, 21, or 35 days), two dietary glucomannan addition levels (0 or 0.1% EGM), and control diet, totaling seven treatments. The following diets were fed: 1) Control diet, 2) pre-starter diet containing FB1, 3) pre-starter diet containing FB1 and 0.1% EGM, 4) starter diet containing FB1, 5) starter diet containing FB1 and 0.1% EGM, 6) grower diet containing FB1, and 7) grower diet containing FB1 and 0.1% EGM. On d 7, broilers fed FB1 presented lower body weight gain and feed intake (p<0.05) compared with control treatment. On d 21, no significant performance differences were detected among treatment groups (p>0.05). At 35 days of exposure to FB1 body weight gain was reduced (p<0.05) compared with broilers fed fumonisin B1 for seven days. From 4 to 7 days and 18 to 21 days of age, FB1 reduced nutrient metabolizability (p<0.05). From 36 to 39 days of age, the EGM allowed maintaining apparent metabolizability for ether extract. It was concluded that the EGM did not reduce FB1 effects on performance or nutrient metabolizability in broilers, except for apparent metabolizability of ether extract.
Abstract in English:ABSTRACT A photonic imaging method that gives the possibility to measure egg quality was applied. Since the method is non-contact and non-destructive we believe that this photonic imaging method may be successfully integrated in the automated inspection systems in the poultry industry. The method involves scanning an invisible infrared light beam over the eggshell, allowing to detect possible cracks and reveal information about the structure of the eggshell. The high resolution, high quality measurements obtained through optical coherence tomography (OCT) make it feasible to be utilized as part of an automated inspection system. In this paper we present an OCT scan image of the egg tip and reconstructed volumetric images of the eggshell surface. The method enables the detection of small cracks on eggs and reveals the detailed inner structures of the cracks.
Abstract in English:ABSTRACT Vitamin D is added to broiler diets to supply its physiological requirement for bone formation. The fast growth rate of modern broilers is often associated with poor bone formation. Increasing vitamin D supplementation levels and the use of more available sources have applied to try to prevent leg problems, to increase carcass yield, and to improve the performance of broilers. The present study evaluated three vitamin D supplementation levels (1) 3,500 IU (control); (2) control + 1,954 IU of 25-hydroxycholecalciferol; and (3) control + 3,500 IU of vitamin D in broiler diets supplied up to 21 days of age. The objective was to investigate if the vitamin D levels above the recommendations could reduce leg problems in broilers. In this experiment, a total of 1,296 one-day-old male and female Cobb(r) 500 broilers were used. A 2 x 3 factorial arrangement was applied, consisting of two sexes and three vitamin D levels. No difference was found between the levels of vitamin D (p > 0.05), the performance of males or females, the gait score, the valgus and varus incidence, the tibial dyschondroplasia incidence, the occurance of femoral degeneration, the bone colorimetric, and the carcass yield. Parts yield differences were found (p > 0.05), except for liver and intestine yields. We concluded that the lowest tested vitamin D level (3,500 IU per kilogram of feed) added to the diet was the best choice in terms of cost/benefit to help minimizing leg problems in broilers.
Abstract in English:ABSTRACT This study aimed at comparing the growth and mineralization of the femur, tibia, and metatarsus of male and female broiler chicks. On the day of hatch, 100 male and 100 female Ross 308 broiler chicks were transferred stainless cages with 10 birds per cage. On d 7, 14, 21, 28, 35, and 42, five males and five females were sacrificed and their femur, tibia, and metatarsus were collected. Results showed that the tibia was the heaviest and the longest and contained the highest content of ash and calcium (Ca) among the three leg bones. The femur had the greatest diameter. The weight, length, diameter, and ash weight of the femur, tibia, and metatarsus linearly increased with age. The ash, Ca, and phosphorus (P) content in the femur and the tibia quadratically increased with age; by contrast, these parameters in the metatarsus linearly increased with age. The bones grew faster in 1 to 21 d of age. The weight, diameter, and ash weight of the three bones of males were higher than those of females. The Ca to P ratio of the three bones (femur, tibia, and metatarsus) was approximately 2.0:1. These data indicate that there are differences in bone growth and mineralization among the femur, tibia, and metatarsus of male or female broiler chicks.
Abstract in English:ABSTRACT The objective of this study was to compare the effects of two methods (electrical water bath or carbonic gas atmosphere) for stunning or killing broiler chickens prior to bleeding on weight loss due to bleeding and meat traits. A completely randomized design with 2 x 2 factorial arrangement (electrical or gas system x stunning or killing) was applied. The time required for stunning and killing and the birds' behavior were evaluated for the gas exposure method. The birds killed by the electrical method retained more blood than those killed by the exposure to gas and, therefore, presented redder meat. The exposure to 10 to 15% CO2 atmosphere caused discomfort reactions in 100% of birds, and the intensity of reaction presented a wide variation: 35% for weak reaction, 40% for intermediate reaction, and 25% for strong reaction. The time required to stun or kill the birds by CO2 exposure varied from 28 to 97 seconds and from 42 to 158 seconds, respectively. It was concluded that the time required to stun or to kill the birds by CO2 exposure widely varied and caused discomfort reactions; however, meat traits were not influenced by the methods used in this experiment.
Abstract in English:ABSTRACT This study was conducted to determine the growth performance, organ weights, and selected blood parameters of replacement laying pullets (development phase) fed increasing levels of wheat bran (WB). A total of 240 70-d-old White Leghorn pullets (Hybrid L33) were evaluated for seven weeks. Birds were assigned to three dietary treatments according to a completely randomized design. Treatments consisted of diets containing 100 (T1), 150 (T2), or 200 g/kg (T3) of WB in partial replacement of corn, with 10 replicates per treatment of eight birds per replicate. Birds fed T2 presented higher body weight (p<0.05) compared with T1 and T3 (1112.52 to 1163.35 g). Also, T2 birds presented higher methionine plus cystine intake relative to T1 (0.38 to 0.40 g/bird/day). Likewise, a higher inclusion of WB (200 g/kg) increased crude fiber (2.29 to 2.63 g/bird/day) and crude fat (1.98 to 3.58 g/bird/day) intakes (p<0.05). However, the experimental treatments did not affect the relative weight of the organs or small intestine and cecum length (p>0.05). Serum concentration of triacylglycerols, cholesterol, calcium, phosphorus, hematocrit, or hemoglobin levels were not significantly different (p>0.05) among treatments. These findings indicate a beneficial effect of use of 150 g/kg of wheat bran on the growth performance of pullets during the development phase; however, the inclusion of this cereal up to 200 g/kg had no effect on organ weights and blood parameters.
Abstract in English:ABSTRACT The objective of this study was to evaluate the effects of 1% dietary glutamic acid on the body weight, intestinal morphometry, and anti-Newcastle antibody titers of broiler chickens submitted to heat stress. One-d-old male broiler chicks (n=120) were distributed according to a 2 x 2 factorial design with two environmental temperatures (thermoneutral or heat stress) and two diets (with 0 or 1% glutamic acid). Heat stress temperature was constantly maintained (24h/day) 5 ºC higher than the thermoneutral temperature. Diets supplied the nutritional requirements of broilers in the pre-starter (1 to 7d) and starter (8 to 21d) phases. Birds were vaccinated against Newcastle disease on d 7 via eye drop. On days 5, 10, 15, and 20, individual body weight was determined, serum samples were collected from five birds, and duodenum samples were collected from four birds per treatment. Serum anti-Newcastle antibody titers were determined by enzyme immunoassay and transformed into log10. Villus height, crypt depth, and villus: crypt ratio were measured in the duodenum. Data were analyzed by ANOVA. Chronic heat stress negatively affected body weight and intestinal morphometry during the pre-starter and starter phases, but had no effect on antibody titers. Dietary glutamic acid supplementation (1%) improved body weight and intestinal integrity of birds submitted to heat stress when compared with non-supplemented and heat-stressed birds.
Abstract in English:ABSTRACT The objective of this experiment was to evaluate the performance of 64- to 79-wk-old laying hens fed diets supplemented with an enzyme complex (EC) and containing increasing sunflower meal (SFM) levels. A total of 384 Hy-Line Brown layers were distributed according to a randomized block design in a 4 × 2 factorial arrangement (four levels of SFM, and inclusion or not of EC), with eight replicates of six birds each unit. The levels of SFM inclusion were 0, 8, 16 and 24%, utilized in two distinct diets. Diets were calculated to meet all the nutritional requirements of birds, except for the nutrients that would be made available by the nutritional matrix of the enzyme complex, with or without utilization of EC. The parameters analyzed were feed intake (g/bird/day), egg production (%/bird/day), egg weight, egg mass (g/bird/day), feed conversion ratio per egg mass, feed conversion ratio per dozen eggs, body weight gain, egg components (yolk, albumen and eggshell) and the economic efficiency index (EEI). There was no interaction between EC addition and the SFM levels in the diet. The addition of EC in the diets of laying hens did not affect egg productive or components parameters. The increase in the SFM levels in the diet presented quadratic effect on egg production and feed conversion ratio per dozen eggs, with calculated optimal sunflower meal inclusion levels of 6.72% and 5.83%, respectively, for each parameter. The best economic efficiency per dozen eggs was obtained with the diet with 16.0% SFM and EC inclusion, whereas per egg mass with the diet with of 24.0% SFM and no EC addition.
Abstract in English:ABSTRACT This experiment was conducted to evaluate the combination effect of low dietary non-phytate phosphorus (NPP) concentrations, phytase (PHY) levels, and 25-hydroxycholecalciferol (25-OH-D3) levels on the growth performance and meat quality of broilers. Two levels of NPP, two levels of PHY, and two levels of 25-OH-D3 resulted in a 2�2�2 factorial arrangements, with eight treatments (TRT). The birds on TRT 1-4 were fed diet 1 (NRC NPP was reduced by 0.1) and the birds on TRT 5-8 were fed with diet 2 (NRC NPP was reduced by 0.2). Each diet was mixed with different levels PHY and 25-OH-D3. Performance and meat quality parameters were measured. Results showed that during entire experiment the most advantageous effects were obtained with TRT 3 (NRC NPP reduced by 0.1 + 600 U/kg phytase + 34.5�g/kg 25-OH-D3) and TRT 4 (NRC NPP reduced by 0.1 + 600 U/kg phytase + 69�g/kg 25-OH-D3). The lowest body weight gain (BWG) and feed intake(FI) were observed with TRT 5 (NRC NPP reduced by 0.2 + 300 U/kg phytase + 34.5�g/kg 25-OH-D3). Lowering NRC NPP by 0.1 to 0.2 significantly reduced weight gain (WG) (p< 0.05) and FI (p< 0.05) during the starter phase (ST), while during grower phase (GF) lowering NRC NPP by 0.1 to 0.2 did not affect WG (p>0.05) and produced small decrease in FI. BWG, FI and feed conversion ratio were not influenced (p>0.05) by different PHY or 25-OH-D3 levels. In addition, the meat color, pH, and shear force were not affected by the different NPP, PHY or 25-OH-D3levels.
Abstract in English:ABSTRACT The digestible lysine requirement for the growth performance of broilers fed diets with decreasing energy to protein ratio and reared under subtropical summer conditions were determined. Treatments consisted of five wheat-based diets with five levels (0.90, 0.95, 1.00, 1.05 and 1.10%) of digestible lysine (DL) and lowering energy to protein ratio (EPR; 13.6, 12.8, 12.0, 11.4 and 11.0), with five replicates of 80 birds each (n = 2000). Body weight gain (BWG) significantly increased by increasing DL levels during wk 1 (p<0.001), while 1.05% DL promoted the highest BWG during the whole experimental period (p<0.01). Increasing DL level resulted in significant reduction in feed intake (FI) and BWG to DL intake ratio (BWG:DLI) during wk 1-3 (p<0.05). Feed:gain ratio (FGR) significantly (p<0.01) improved with DL levels up to 1.05% during the whole experimental period, except for wk 5. The increasing ambient temperature had a linear effect on the all growth parameters. It is inferred that best growth performance can be obtained at 1.05% DL, maintaining all other AA constant under subtropical summer conditions.
Abstract in English:ABSTRACT The aim of the current study was to determine the effects of low (33.3-36.7 ˚C), control (37.8-38.2 ˚C) and high (38.9-40.0 ˚C) eggshell temperatures (ESTs) between days 10-18 of incubation on broiler performance and susceptibility to ascites. The effects of different ESTs on hatchling weight were significant. The highest residual yolk sac weight (7.7 g) and relative yolk sac weight (18.7%) were observed in high EST group, whereas the highest yolk-free body weight (36.22 g) was obtained in the control group. Relative post-hatch heart weight was lower in the high EST group (0.64%) compared with the control (0.84%) and low (0.88%) EST groups. At 6 weeks of age, body weights of broilers of the low, control, and high EST groups were 2172.6, 2543.9, 2282.6 g respectively. During the 6-week life span, the feed conversion ratio of broilers in the low (1.86) and high (1.83) EST groups was significantly worse than of those in the control EST group (1.68). Higher packed cell volume (PCV) and red blood cell (RBC) values were obtained in the high EST group relative to the other groups, whereas hemoglobin (Hb) level was lower in the control group than in the others. The relative heart weights and right ventricular to total ventricular (RV:TV) ratio for low, control, and high EST group were 0.46, 0.40, 0.37 g and 0.27, 0.28, 0.33% respectively. In conclusion, incubation length, yolk sac absorption, residual yolk sac weight, yolk-free BW, post-hatch performance, and heart, blood parameters, abdominal fluid accumulation at slaughter age are affected by changes in EST between incubation days 10 to 18.
Abstract in English:ABSTRACT Pendulous crop is a physiological disorder, which etiology is still unknown and it is characterized by abnormal dilation of the crop of poultry. This article aims at reporting a case of high incidence of pendulous crop in male and female broilers Cobb 500, as well as to discuss its possible causes and consequences. In an experiment with broilers performed at the experimental facilities of Laboratório de Ensino Zootécnico of UFRGS, a high incidence (9.5%) of pendulous crop was observed. Genetic predisposition is the most frequently documented and accepted cause of that condition. Despite presenting the same live weight as normal broilers, birds with pendulous crop had lower carcass weight due to dehydration and malnourishment, and should be culled after diagnosis. Therefore, further studies on the origin and control of this physiological disorder are warranted.
Abstract in English:ABSTRACT Avian encephalomyelitis is caused by an Hepatovirus and primarily affects chickens. Chickens of all ages are susceptible to the virus, but the nervous symptoms are manifested only in young chicks, between one to five weeks of age. During the last thirty years, avian encephalomyelitis appeared to be well controlled by breeder vaccination. However, the increase of the number of cases is causing concern in the poultry industry. In the present study, we performed a retrospective analysis of the cases presenting histological lesions compatible with avian encephalomyelitis in broilers. The evaluated cases affected broilers from one to 35 days old from the southern region of Brazil. Only cases with compatible microscopic lesions and associated with clinical symptoms in the field were considered. In addition the histopathological diagnosis, sera were tested by ELISA (Enzyme-Linked Immunosorbent Assay). Considering the clinical, histopathological, and serological evidences, the disease was confirmed, showing an increase in outbreaks from the last quarter of 2012, extending through 2013. The cause of this increase is not clear, although we suspect vaccine or vaccination error. Enhancing vaccinated breeders flocks monitoring before the beginning of egg production and/or using a protocol with two vaccinations is recommended.