Production of fumonisins byFusarium species of Taiwan

Twenty-nineFusarium species isolated from various sources in different districts of Taiwan were tested for their ability to produce fumonisins in corn cultures. OnlyFusarium moniliforme produced fumonisin B₁ (FB₁) and fumonisin B₂ (FB₂). The finding that the other 28Fusarium species produced neither FB₁ nor FB₂ is preliminary because only one strain per species was studied. The detection of FB₁ and FB₂ in cultures ofF. moniliforme was demonstrated by TLC and HPLC, and FB₁ was further confirmed by mass spectrometry. In a separate experiment, in which 38 strains ofF. moniliforme were tested for fumonisins, approximately 66% (25/38) produced FB₁ and/or FB₂. Of the 25 strains, 14 produced only FB₁ and 11 produced both FB₁ and FB₂, and the amounts of FB₁ and FB₂ produced by different strains varied greatly. This is the first report that fumonisins are found in corn cultures experimentally infected withF. moniliforme strains from Taiwan. It is safe to assume that fumonisin producing strains ofF. moniliforme are widely distributed among the economic crops such as corn, rice, sugarcane, and sorghum throughout the Island.Abbreviations FB₁ Fumonisin B₁ - FB₂ Fumonisin B₂ - OPA o-phthalidialdehyde     

Production of zearalenone,moniliformin and trichothecenes in intact sugar beets under laboratory conditions

Five toxigenic isolates of Fusarium species were tested for the production of zearalenone, moniliformin and trichothecenes (deoxynivalenol, 15-acetyldeoxynivalenol, T-2, HT-2 and neosolaniol) when grown on solid sugar beet slices in the laboratory for thirty days. The isolates were also grown on a solid rice medium for comparison. High zearalenone and trichothecene-producing isolates originally obtained from corn and corn-based feedstuff were compared with isolates obtained from sugar beets. One moniliformin-producing isolate from wheat was included in the study. With the exception of moniliformin, all toxins were produced on both substrates; however, the rice medium yielded the greater concentrations except for HT-2 which was produced on sugar beets in equal or greater concentrations. Zearalenone production on rice reached 729–1943 gmg/g whereas on sugar beet it reached 72–193 gmg/g. The moniliformin-producing isolate grew well on both substrates; however, moniliformin was produced only on the rice substrate. This study demonstrates for the first time that Fusarium species can produce both zearalenone and the trichothecenes on a sugar beet substrate.     

Identification of deoxynivalenol, 3-acetyldeoxynivalenol and zearalenone in the galactose oxidase-producing fungus Dactylium dendroides

The galactose oxidase-producing fungus Dactylium dendroides was re-identified as a Fusarium species. Fungi of this genus are well known for the production of mycotoxins. Verification of growth of this fungus on rice, corn and liquid medium described for the production of galactose oxidase is provided to determine whether the fungus could produce Fusarium toxins, namely, moniliformin, fusaric acid, fumonisin, zearalenone and the trichothecenes, deoxynivalenol, 3-acetyldeoxynivalenol, fusarenone, nivalenol, diacetoxyscirpenol, neosolaniol, and toxin T-2. Under the culture conditions used, deoxynivalenol, 3-acetyldeoxynivalenol and zearalenone were detected in the fungal culture medium. The finding is consistent with the hypothesis that the fungus is in fact a Fusarium species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Prevalence of Fusarium species of the Liseola section on selected Colombian animal feedstuffs and their ability to produce fumonisins

A total of 57 samples of feedstuffs commonly used for animal nutrition in Colombia (corn, soybean, sorghum, cottonseed meal, sunflower seed meal, wheat middlings and rice) were analyzed for Fusarium contamination. Fusarium fungi were identified at species level by means of conventional methods and the ability to produce fumonisins of the most prevailing species was determined. A total of 41 of the feedstuffs analyzed (71.9%) were found to contain Fusarium spp. Most contaminated substrates were corn (100%), cottonseed meal (100%), sorghum (80%), and soybean (80%). Wheat middlings and rice showed lower levels of contamination (40% and 20%, respectively), while no Fusarium spp. could be isolated from sunflower seed meal. The most prevalent species of Fusarium isolated were F. verticilliodes (70.8%), F.␣proliferatum (25.0%), and F. subglutinans (4.2%). All of them correspond to section Liseola.Production of fumonisins on corn by the isolated Fusarium was screened through liquid chromatography. Almost all strains of F. verticilliodes (97.1%) produced FB1 (5.6–25,846.4 mg/kg) and FB2 (3.4–7507.5 mg/kg). Similarly, almost all strains of F.␣proliferatum (91.7%) produced fumonisins but at lower levels than F.␣verticilliodes (FB1 from 6.9 to 3885.0 mg/kg, and FB2 from 34.3 to 373.8 mg/kg), while F. subglutinans did not produce these toxins. This is the first study in Colombia describing toxigenic Fusarium isolates from␣animal feedstuffs.     

Genetic and mutational tools for investigating the genetics and molecular biology of trichothecene production inGibberella pulicaris (Fusarium sambucinum)

Gibberella pulicaris (Fusarium sambucinum) is a promising organism for studying the genetics and regulation of trichothecene biosynthesis; conditions for obtaining fertile crosses have been defined (Desjardins & Beremand, 1987) and crosses between natural variants have provided some information about the number, location, arrangement, and role of genes which determine trichothecene production (Desjardins & Beremand, 1987; Beremand & Desjardins, 1988). The development of some additional experimental tools and methodologies required for the further genetic analysis of trichothecene production inG. pulicaris are described in the present study. A highly fertile, isogenic line was constructed forG. pulicaris strain R-6380. The ability to readily generate mutants in this strain was also demonstrated. Both biochemical and morphological mutants were obtained following UV-mutagenesis. The inheritance of some of these mutations through meiosis indicated that they will be useful genetic markers for crosses and mapping studies. Since strain R-6380 is also transformable (Salch & Beremand, 1988), it is an excellent choice for transmission and molecular genetic studies involving trichothecene production.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Vitamin e hypervitaminosis in laying hens

The aim of this investigation was to contribute to the problem of overdosing vitamin E. A total of 80 laying hens, divided into 5 groups, were fed diets supplemented with 0; 100, 1000, 10 000 and 20 000 mg/kg vitamin E over a period of 20 weeks within two brooding tests. Laying performance and hatching parameters were registered. All vitamin E doses did not significantly influence health and performances of hens. Vitamin E content of eggs increased from 1 to 4, 21, 46 and 51 mg per egg with vitamin E supplementation. High doses decreased oxidative stability of abdominal fat, vitamin A concentration of liver and egg yolk colour. In both tests vitamin E supplements of 10 000 and 20 000 mg/kg feed resulted in a decrease of living hatched chicken, which demonstrated an adverse effect. Further studies seem to be necessary to explain the effect.     

Occurrence and variability of mycotoxigenicFusarium species associated to wheat and maize in the South West of Spain

The contamination of cereals with mycotoxins produced by species ofFusarium is an important risk to human and animal health. The toxigenic profile is different depending on theFusarium species considered and, in some species, differences can also be observed at intraspecific level. Information about the distribution and variability of the mycotoxigenicFusarium species allow prediction of the toxins that may occur and to devise control strategies. In this work, the occurrence of mycotoxigenicFusarium species associated to cereals was analysed in a wide sample of durum wheat fields (Triticum durum Desf.) and maize from the South West of Spain (Andalucía).F. equiseti, F. graminearum andF. culmorum were the most frequentFusarium species detected in wheat fields followed byF. sambucinum andF. avenaceum, whereas in the case of maize,F. verticillioides andF. proliferatum were the onlyFusarium species present. The relationships of the Spanish isolates from theF. equiseti, F. avenaceum andF. sambucinum species were analysed by nucleotide sequence comparison of a partial region of the Elongation Factor 1 alpha (EF-1α) with other sequences available in data bases. The results indicated thatF. avenaceum andF. equiseti showed high variability and that the SpanishF. equiseti isolates seemed to belong toF. equiseti type II. Presented at the EU-USA Bilateral Workshop on Toxigenic Fungi & Mycotoxins, New Orleans, USA, July 5–7, 2005 Financial support: MCYT (AGL2004/07549/C05/5). M. Jurado was supported by pre-doctoral fellowship by the MCYT     

Development of locomotion over inclined surfaces in laying hens

The purpose of the present study was to evaluate locomotor strategies during development in domestic chickens (Gallus gallus domesticus); we were motivated, in part, by current efforts to improve the design of housing systems for laying hens which aim to reduce injury and over-exertion. Using four strains of laying hens (Lohmann Brown, Lohmann LSL lite, Dekalb White and Hyline Brown) throughout this longitudinal study, we investigated their locomotor style and climbing capacity in relation to the degree (0 to 70°) of incline, age (2 to 36 weeks) and the surface substrate (sandpaper or wire grid). Chicks and adult fowl performed only walking behavior to climb inclines ⩽40° and performed a combination of wing-assisted incline running (WAIR) or aerial ascent on steeper inclines. Fewer birds used their wings to aid their hind limbs when climbing 50° inclines on wire grid surface compared with sandpaper. The steepness of angle achieved during WAIR and the tendency to fly instead of using WAIR increased with increasing age and experience. White-feathered strains performed more wing-associated locomotor behavior compared with brown-feathered strains. A subset of birds was never able to climb incline angles >40° even when using WAIR. Therefore, we suggest that inclines of up to 40° should be provided for hens in three-dimensional housing systems, which are easily negotiated (without wing use) by chicks and adult fowl.     

云南蛋鸡源肠炎沙门氏菌的分离鉴定及生物学特性

[背景]肠炎沙门氏菌(Salmonella enteritidis)是一种重要的人畜共患病原菌,禽类的肉制品及蛋类是其重要的传播途径.[目的]确诊云南某蛋鸡场疑似沙门氏菌感染病原情况.[方法]无菌采集发病蛋鸡肝脏组织进行细菌分离培养鉴定,对获得的菌株进行耐药性分析、致病性实验及毒力基因的检测.[结果]鉴定该分离菌为肠炎...     

Ochratoxin A and fumonisins (B1 and B2) in maize from Balkan nephropathy endemic and non endemic areas of Croatia

The occurrence of ochratoxin A, fumonisin B1 and B2 has been investigated in maize samples collected in 1996 (105 samples) and 1997 (104 samples) in 14 counties of Croatia, including Brodsko-Posavska county, the main area of Balkan endemic nephropathy in Croatia. Ochratoxin A and fumonisins co-occurred in 21% of the examined samples. In particular, ochratoxin A (OTA) was found in 10 samples (10%) of the 1996 and 36 samples (35%) of the 1997 crops with mean concentrations of positive samples of 37.9 ng/g and 57.1 ng/g, and highest concentrations at 223.6 ng/g and 613.7 ng/g, respectively. Similar incidence of OTA contamination was observed in 1996 samples from both endemic and non endemic areas of Balkan nephropathy, whereas a significant difference (P<0.01) was found between the two areas in 1997, with 50% and 20% incidence of contamination in the endemic and non endemic area, respectively, and relevant OTA mean concentration of positive samples of 73.4 ng/g and 20.2 ng/g. High incidence of infection byPenicillium spp. (potential OTA producers) was found in all tested samples, with mean values of 88% and 93% in samples of 1996 and 1997, respectively. With respect to fumonisin B1 (FB1) and B2 (FB2) all but one of the 1996 samples were contaminated, with highest and mean concentrations of positive samples (FB1+FB2) at 11661 ng/g and 645 ng/g, respectively. Similar incidence of positive samples (93%), but lower contamination levels (mean 134 ng/g, maximum 2524 ng/g) were found in 1997 samples. The results of fumonisin analysis were in agreement with the mycological analysis showing higher incidence of Fusarium infection in samples of 1996 with respect to those of 1997. These data provide additional information on the occurrence of ochratoxin A in Balkan endemic nephropathy areas and, for the first time, its co-occurrence with other nephrotoxic compounds, such as fumonisins, that may contribute to the disease development. However the finding of these mycotoxins in the non-endemic areas, also at high levels, do not allow to draw a conclusion about their role in the etiology of the disease.     

Comparison of in vitro cytotoxicity of Fusarium mycotoxins,deoxynivalenol, T-2 toxin and zearalenone on selected human epithelial cell lines

Three human epithelial cell lines (CaCo-2, HEp-2 and HeLa) implicated as potential targets for three Fusarium toxins were tested for the extent of survival on exposure to increasing toxin concentration and incubation periods. Cytotoxicity assay using 3(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) was carried out with deoxynivalenol (DON), T-2 toxins and zearalenone (ZON) on CaCo-2, HEp-2 and HeLa cell lines. Of the three cell lines used, HeLa was the most sensitive, eliciting cell death after 2 days exposure at 100 ng ml^–1with T-2 toxin. HeLa was the only cell line to exhibit cytotoxicity towards ZON showing cell death at 1000 ng ml^–1after 2 days which increased to 4 days, showing substantial cell death at 200 ng ml^–1. HEp-2 was sensitive to DON showing cell death after 2 days (100 ng ml^–1) with complete cell death occurring at 200 ng ml^–1 after 4 days of exposure. Substantial cytoxicity of T-2 towards HEp-2 occurred after 2 days at 1000 ng ml^–1 and complete cell death occurred with 100 ng ml^–1 at day 4. The CaCo-2 cell line was generally resistant to the mycotoxins tested between 100 and 1000 ng ml^–1. This study shows that cytotoxicity of Fusarium toxins to epithelium cell lines is concentration- and time- dependant and results from ZON–HeLa interaction indicate possible cell type-mycotoxin specificity.     

Cytotoxicity of Fusarium species mycotoxins and culture filtrates of Fusarium species isolated from the medicinal plant Tribulus terrestris to mammalian cells

Ayurvedic medicine, which uses decoctions made of medicinal plants, is used to cure diseases in many Asian countries including Sri Lanka. Although proper storage facilities for medicinal plants are unavailable in Sri Lanka, neither the potential for growth of toxigenic fungi nor their ability to produce mycotoxins in stored medicinal plants has been investigated. We isolated three Fusarium species, F. culmorum, F. acuminatum and F. graminearum from the medicinal plant Tribulus terrestris. Culture extracts of the 3 Fusarium spp. were cytotoxic to mammalian cell lines BHK-21 and HEP-2. Three toxic metabolites produced by Fusarium spp; T-2 toxin, zearalenone, and diacetoxyscirpenol were also cytotoxic to the same mammalian cell lines. The 3 Fusarium spp. grown on rice media produced zearalenone. Plant material destined for medicinal use should be stored under suitable conditions to prevent growth of naturally occurring toxigenic fungi prior to its use.     

Oviduct-specific expression of tissue plasminogen activator in laying hens

Egg-laying hens are important candidate bioreactors for pharmaceutical protein production because of the amenability of their eggs for protein expression. In this study, we constructed an oviduct-specific vector containing tissue plasminogen activator (tPA) protein and green fluorescent protein (pL-2.8OVtPAGFP) and assessed its expression in vitro and in vivo. Oviduct epithelial and 3T3 cells were cultured and transfected with pL-2.8OVtPAGFP and pEGP-N1 (control vector), respectively. The pL-2.8OVtPAGFP vector was administered to laying hens via a wing vein and their eggs and tissues were examined for tPA expression. The oviduct-specific vector pL-2.8OVtPAGFP was expressed only in oviduct epithelial cells whereas pEGP-N1 was detected in oviduct epithelial and 3T3 cells. Western blotting detected a 89 kDa band corresponding to tPA in egg white and oviduct epithelial cells, thus confirming expression of the protein. The amount of tPAGFP in eggs ranged 9 to 41 ng/mL on the third day after vector injection. The tPA expressed in egg white and oviduct epithelial cells showed fibrinolytic activity, indicating that the protein was expressed in active form. GFP was observed only in oviducts, with no detection in heart, muscle, liver and intestine. This is the first study to report the expression of tPA in egg white and oviduct epithelial cells using an oviduct-specific vector.     

Modulation of energy and protein supplies in sequential feeding in laying hens

Sequential feeding (SF) consists of splitting energy (E) and protein/calcium (P) fractions temporally, improving the feed conversion ratio (FCR) of hens compared with a continuous distribution during the day. In a previous study, the E fraction (with a low level of protein) was provided in the morning, whereas the P fraction (with low level of energy) was given in the afternoon. However, there is no clear evidence that a requirement in energy or proteins is connected to these distribution sequences, whereas the requirement for calcium is known to be required in the afternoon. To evaluate the effects on performances of the modulation of energy and protein supplies in SF, five different sequential treatments were offered: E0P0/E0P0; E+P+/E−P−; E+P−/E−P+; E0P+/E0P− and E+P0/E−P0 where E+ represents a high energy level, E0 a moderate one and E− a low one (with the same meaning for P regarding protein supply). Afternoon fractions were provided with particulate calcium. A total of 168 Hendrix hens were housed in individual cages from 20 to 39 weeks of age in two environmentally contrasted rooms. Feed intake in the morning and afternoon fractions, egg production, egg weight, BW and weight of digestive organs were recorded. No diet effect was observed concerning feed intake, egg production and BW. These results suggested that hens are not able to fit their feed intake on energy or protein level of fractions within half-day duration, whereas at the day scale same protein and energy intakes were observed. Moreover, the time of nutrient distribution in feeding did not seem to have an impact on birds’ performances. These studies have also demonstrated that, despite strong environmental pressure, the hens with SF had attenuated performance but continue to produce eggs.     

The presence of extreme feather peckers in groups of laying hens

Feather pecking is a serious economic and welfare problem in laying hens. Feather damage occurs mainly through severe feather pecking (SFP). Selection experiments have proved that this behavior is heritable and lines have been divergently selected for high (HFP) and low feather pecking (LFP). The number of bouts of SFP per hen follows a Poisson distribution with a maximum nearby 0. A few studies indicate that the distribution within flocks is not homogenous but contains sub-groups of birds showing extremely high levels of feather pecking (EFP). It was the aim of the current study to re-analyze data on SFP of lines selected for HFP/LFP and their F2 cross so as to uncover hidden sub-populations of EFP birds. Data of seven selection generations of HFP and LFP selection lines as well as their F2 cross have been used. We fitted a two-component mixture of Poisson distributions in order to separate the sub-group of EFP from the remaining birds. HFP and LFP lines differed mainly in mean bouts per bird. The proportion of EFP was only marginal in the LFP as compared with the HFP and the F2 population. Selection for LFP did not result in total elimination of EFP. The presence of even small proportions of EFP may play an important role in initiating outbreaks of feather pecking in large flocks. Further studies on feather pecking should pay special attention to the occurrence of EFP sub-groups.     

Cytotoxic and immunotoxic effects of Fusarium mycotoxins using a rapid colorimetric bioassay

A colorimetric MTT (tetrazolium salt) cleavage test was used to evaluate cytotoxicity of twenty-three Fusarium mycotoxins on two cultured human cell lines (K-562 and MIN-GL1) as well as their inhibitory effect on proliferation of phytohemagglutinin-stimulated human peripheral blood lymphocytes. The values of 50% inhibition of lymphocyte blastogenesis were very close to the 50% cytotoxic doses observed with the more sensitive cell line (MIN-GL1). T-2 toxin was the most cytotoxic with CD₅₀ and ID₅₀ values less than 1 ng/ml. Type A trichothecenes were the most cytotoxic followed by the type B trichothecenes; the non-trichothecenes were the least cytotoxic. The MTT cleavage test, in conjunction with cell culture, is a simple and rapid bioassay to evaluate cytotoxicity and immunotoxicity of Fusarium mycotoxins.Abbreviations Ac acetyl - ACU acuminatin - DAS diacetoxyscirpenol - DON deoxynivalenol - FUS fusarenon-X - HT-2 HT-2 toxin - MC mononuclear cell - MTT 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide - NEO neosolaniol - NIV nivalenol - NT-1 4,8-diacetoxy T-2 tetraol - PBS phosphate buffered saline - TAT-2T tetraacetoxy T-2 tetraol - T-2 T-2 toxin     

Occurrence ofAlternaria andFusarium mycotoxins in winter wheat from domestic crop in year 2003

The aim of this study was a monitoring of the occurrence ofAlternaria andFusarium mycotoxins in winter wheat from domestic crop in the year 2003. Altenuene was determined in 56 (100%) samples of winter wheat, range 14.5–41 μg/kg, mean 25 μg/kg. Alternariol was determined in 16 (28.6%) samples of winter wheat, range 6.3–22.1 μg/kg, mean 5.7 μ/kg. DON was determined in 42 (100%) samples of winter wheat, range 250–3500 μg/kg, mean 330 μg/kg. T2-toxin was determined in 42 (100%) samples of winter wheat, range 25–337 μg/kg, mean 99 μg/kg. ZEA was not determined in samples of winter wheat. Presented at the 26^th Mykotoxin-Workshop in Herrsching, Germary, May 17–19, 2004 Financial support. Supported (one part of experiments, the determination of Fusarium mycotoxins) by the Ministry of Agricu ture of the Czech Rebublic (Propect No QF3121)     

Circadian rhythms and dynamic dietary calcium feeding affect laying performance,calcium and phosphorus levels in laying hens

This study was conducted to determine the effects of dynamic feeding low and high Ca in layer diets on laying performance, calcium, and phosphate concentration in both serum and tibia. 180 Brown Hy-line, 41 weeks old of laying hens were randomly distributed into 3 groups with 6 replicates and each replicate was 10 birds. During the 10-week experiment period, control group (CON group) received control diet (contained 3.4% Ca) at both 0730 and 1530 h, low–high group (LH group) received low Ca diet (contained 3.2% Ca) at 0730 h and high Ca diet (contained 3.6% Ca) at 1530 h, while high–low group (HL group) received the contrary to the LH group. Blood and tibia samples were collected at 4 h intervals. The dynamic Ca feeding schedule exerted significant effects on Ca intake in the morning and afternoon, also on feed conversion ratio (FCR) of the HL group comparing to the LH group (p<0.05). Serum Ca in the LH group were lower at 0000, 0400 and 1600 h comparing with the CON group (p < 0.05), and serum Ca: P ratio values in the LH group were lower at 0000, 0800, and 2000 h comparing with the CON group (p<0.05). These results indicated that dynamic feeding Ca improved FCR and affected the circadian variation of serum Ca concentration in laying hens.     

Effects of probiotic supplementation on performance traits,bone mineralization,cecal microbial composition,cytokines and corticosterone in laying hens

Recent researches have showed that probiotics promote bone health in humans and rodents. The objective of this study was to determine if probiotics have the similar effects in laying hens. Ninety-six 60-week-old White Leghorn hens were assigned to four-hen cages based on their BW. The cages were randomly assigned to 1 of 4 treatments: a layer diet mixed with a commercial probiotic product (containing Enterococcus faecium, Pediococcus acidilactici, Bifidobacterium animalis and Lactobacillus reuteri) at 0, 0.5, 1.0 or 2.0 g/kg feed (Control, 0.5×, 1.0× and 2.0×) for 7 weeks. Cecal Bifidobacterium spp. counts were higher in all probiotic groups (P<0.001) compared with the control group. The percentage of unmarketable eggs (cracked and shell-less eggs) was decreased in both 0.5× and 2.0× groups compared with the control group (P=0.02), mainly due to the reduction of shell-less eggs (P=0.05). The increases in tibial and femoral mineral density and femoral mineral content (P=0.04, 0.03 and 0.02, respectively), with a concomitant trend of increases in humerus mineral density and tibial mineral content (P=0.07 and 0.08, respectively), occurred in the 2.0× group. However, the bone remodeling indicators of circulating osteocalcin and c-terminal telopeptide of type I collagen were similar among all groups (P>0.05). In addition, the plasma concentrations of cytokines (interleukin-1β, interleukin-6, interleukin-10, interferon-γ and tumor necrosis factor-α) and corticosterone as well as the levels of heterophil to lymphocyte ratio were similar between the 2.0× group and the control group (P>0.05). In line with these findings, no differences of cecal tonsil mRNA expressions of interleukin-1β, interleukin-6 and lipopolysaccharide-induced tumor necrosis factor-α factor were detected between these two groups (P>0.05). These results suggest that immune cytokines and corticosterone may not involve in the probiotic-induced improvement of eggshell quality and bone mineralization in laying hens. In conclusion, the dietary probiotic supplementation altered cecal microbiota composition, resulting in reduced shell-less egg production and improved bone mineralization in laying hens; and the dietary dose of the probiotic up to 2.0× did not cause negative stress reactions in laying hens.     

Alternative methods for disposal of spent laying hens: Evaluation of the efficacy of grinding,mechanical deboning,and of keratinase in the rendering process

Besides the challenges of mortality and litter disposal, the poultry industry must find economical means of disposing of laying hens that have outlived their productive lives. Because spent hens have low market value and disposing of them by composting and burial is often infeasible, finding alternative disposal methods that are environmentally secure is prudent. The feasibility of grinding or mechanically deboning spent hens with and without prior mechanical picking was evaluated for the production of various proteinaceous by-product meals. The end products were analyzed for nutrient content and found to be high in protein (35.3–91.9% CP) and, with the exception of the feathers, high in fat (24.1–58.3%), making them potentially valuable protein and energy sources. After considering physical and economic feasibility, mechanical deboning was determined to be a logical first step for the conversion of spent hens into value-added by-product meals. Because the hard tissue fraction (primarily feathers, bones, and connective tissue) generated by mechanically deboning the hens presents the greatest challenge to their utilization as feedstuffs, attention was focused on technologies that could potentially improve the nutritional value of the hard tissue for use as a ruminant protein source. Traditional hydrolysis of this hard tissue fraction improved its pepsin digestibility from 74% to 85%; however, subsequent keratinase enzyme treatment for 1 h, 2 h, 4 h, or 20 h after steam hydrolysis failed to improve the pepsin or amino acid digestibility any further (P > 0.10). Enzyme hydrolysis did, however, increase the quantities of the more soluble protein fractions (A: 45.5, 46.6, 52.8, 51.6, and 55.8% of CP; B₁: 3.2, 9.8, 6.0, 4.6, and 4.1% of CP; B₂: 11.7, 18.1, 22.8, 29.6, and 22.0% of CP for 0, 1 h, 2 h, 4 h, and 20 h, respectively) and reduced quantities of the less soluble fractions (B₃: 30.2, 18.1, 10.8, 5.5, and 10.2% of CP; C: 9.4, 7.5, 7.6, 8.8, and 7.9% of CP for 0, 1 h, 2 h, 4 h, and 20 h, respectively). The protein digestibility of the steam hydrolyzed hard tissue fraction from the mechanical deboning of spent hens was found to be comparable to the digestibility of feather meal, but post-hydrolysis keratinase treatment did not improve feeding value for ruminants.