PCR-based assay for the rapid detection of fumonisin-producing Fusarium species in maize-based animal and poultry feeds in Karnataka,India

Abstract

One hundred and fourteen animal feedstuffs and eighty poultry feed mixtures commonly used for animal and poultry nutrition in Karnataka, India, were analysed for Fusarium contamination. The total counts of fusaria in animal feeds and poultry feed mixtures revealed a high incidence of F. verticillioides, being isolated from all positive samples. Most contaminated samples were maize pellets (71.4%), cotton seed (66.6%), maize powder (60%) and fine wheat bran (50%), respectively, while no Fusarium species was isolated from Bengal gram husk and wheat flakes. All the Fusarium species were identified by the PCR method using genus specific ITS and group specific FUM 1 primers. Of the 374 Fusarium isolates tested with ITS set of primers, all fusaria scored positive, whereas only 244 (65%) isolates tested positive with the FUM 1 set of primers. The specificity of the primers provides the basis for a simple, accurate and precise detection of Fusarium species that represents fumonisin producers, which are a considerable risk for animal, poultry and human health.     

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.     

Natural Occurrence of 16 Fusarium Toxins in Grains and Feedstuffs of Plant Origin from Germany

A total of 220 samples comprising cereals, cereal byproducts, corn plants and corn silage as well as non-grain based feedstuffs was randomly collected during 2000 and 2001 from sources located in Germany and analysed for 16 Fusarium toxins. The trichothecenes scirpentriol (SCIRP), 15-monoacetoxyscirpenol (MAS), diacetoxyscirpenol (DAS), T-2 tetraol, T-2 triol, HT-2 and T-2 toxin (HT-2, T-2), neosolaniol (NEO), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivealenol (15-ADON), nivalenol (NIV) and fusarenon-X (FUS-X) were determined by gas chromatography/mass spectrometry. Zearalenone (ZEA) and α- and β-zearalenol (α- and β-ZOL) were analysed by high performance liquid chromatography with fluorescence and UV-detection. Detection limits ranged between 1 and 19 μg/kg. Out of 125 samples of a group consisting of wheat, oats, corn, corn byproducts, corn plants and corn silage only two wheat samples did not contain any of the toxins analysed. Based on 125 samples the incidences were at 2–11% for DAS, NEO, T-2 Triol, FUS-X, α- and β-ZOL, at 20–22% for SCIRP, MAS, T-2 tetraol and 3-ADON, at 44–74% for HT-2, T-2, 15-ADON, NIV and ZEA, and at 94% for DON. Mean levels of positive samples were between 6 and 758 μg/kg. Out of 95 samples of a group consisting of hay, lupines, peas, soya meal, rapeseed meal and other oilseed meals, 64 samples were toxin negative. DAS, T-2 triol, NEO and FUS-X were not detected in any sample. The incidences of DON and ZEA were at 14 and 23% respectively, those of the other toxins between 1–4%, mean levels of positive samples were between 5 and 95 μg/kg.     

猪常用植物性饲料氨基酸消化率

采用8头初始体重平均为27.2kg±2.3kg、在回肠末端装有简单T型瘘管的杜长大三元杂交公猪,手术康复后随机分成2组,每组4头,每组采用同一豆粕-玉米淀粉基础日粮,按拉丁方设计,用于测定豆粕、棉粕、菜粕、稻谷、油糠、统糠和玉米7种饲料原料的回肠和粪氨基酸消化率。结果表明,所测氨基酸平均表观消化率回肠比粪低5%~8%(P<0.05);稻谷氨基酸表观消化率比玉米低5%;所测原料氨基酸表观消化率与中国饲料数据库和前人报道结果基本吻合。     

Einfluß der Futterration und des Entnahmezeitpunktes von Pansensaft auf seine Zusammensetzung in Untersuchungen an Kühen

Different starch components were added to rations with maize silage measuring the influence of the supplements on the digestibility and energy content of maize silage in cattle (n = 36) and sheep (n = 36). The starch‐rich components maize, wheat, barley, oat, cassava meal and potatoe starch were added to the total rations on average of 33%. All supplements were ground (sieve of 2.5 mm), additionally, wheat was added in extruded or crushed form. All rations including the control ration without starch components were supplemented with soybean meal to an average crude protein content of 12.4% DM. The mean starch content of the total rations was 42% DM.

The supplementation of maize silage with the different starch components increased the digestibility of the organic matter of all rations on average only slightly from 77% to 78% in cattle, while the mean digestibility in sheep rised from 74% to 79% more pronounced. However, the influence of the different starch components on the digestibility was similiar for both ruminants. The supplementation with the ground, crushed or extruded wheat, with barley and with oats impaired the digestibility of the crude fibre of the total ration and of the maize silage, respectively. Therefore, the digestibility of the organic matter of maize silage was reduced and the energy content decreased by about 6% in comparison to the control ration without starch supplementation. However, the supplementation with maize, cassava meal or pota‐toe starch had no negative effects on the digestiblity and energy content of maize silage.     

Occurrence of cyclopiazonic acid in feeds and feedstuffs in Tamil Nadu,India

Out of 100 samples (67 randomly collected and 33 known to contain aflatoxin) of feeds and feedstuffs examined, 40 showed cyclopiazonic acid (CPA) activity i.e., maize 10/26, groundnut cake 10/20, sunflower seed cake 7/10, sorghum 4/10, wheat 1/1, little millet 1/1, rice 1/1, deoiled rice bran 0/1, fishmeal 0/1, chick mash 1/3, grower mash 0/7, layer mash 3/11, broiler mash 2/6 and cattle feed 0/2, concentrations ranged from 0.4 to 12, 0.5 to 20, 0.3 to 20, 0.3 to 20,20, 10, 10, 1.5, 1 to 15, 8, and 15 ppm, respectively. Co-occurrence of CPA was found in 14 of 33 aflatoxin-containing samples. Part of Ph.D. thesis submitted by the first author to the Tamil Nadu Veterinary and Animal Sciences University, Madras 600 007, India.     

Effect of diets on growth,digestibility, carcass and meat quality characteristics of four rabbit breeds

The current study was conducted to evaluate three diets using four rabbit breeds. A total of 320 male weaned rabbits representing four breeds named V-line, Saudi-1, Saudi-2 and Saudi-3 were randomly distributed into three comparable dietary treatments. Three levels of indigenous feedstuffs (IFS) of 42.5%, 65% or 87.5% (alfalfa hay, barley and wheat bran) were substituted for the same levels of non-indigenous feedstuffs in the diet (corn grain, soybean, molasses, and limestone) to form three diets named D1, D2 and D3, respectively. These dietary treatments were used to evaluate post-weaning growth performance, feed intake, feed conversion, carcass and lean composition, and nutrients digestibility in four rabbit breeds.Partial replacement of IFS with non-indigenous feedstuffs attained significant increase in growth performance. The diet containing 87.5% IFS led to significant increase in daily weight gains by 8.4, 4.0, 8.1, 6.2 and 6.7 g at age intervals of 5–6, 7–8, 9–10, 10–11 and 11–12 weeks compared to the control diet, respectively. The diet containing 87.5% IFS showed a significant reduction in feed conversion ratio compared to control diet during the experimental periods. The diet containing 65% IFS showed significant increases in pre-slaughter weight (112 g) and hot carcass weight (89 g), while dressing percent, offal weight and percent, and non-carcass weights and percentages (head, fur, legs + tail, viscera) were not significantly different. The diet containing 87.5% IFS gave an increase of 1.1%, 6.4%, 8.8%, 17.9%, 7.4%, and 19.6% in digestibility coefficients of organic matter (OM), crude protein (CP), neutral detergent fibre (NFD), acid detergent fiber (ADF), hemi-cellulose (HC) and cellulose (C) compared to control diet, respectively.Feed intakes were moderate and ranged from 69 to 124 g for V-line, 77 to 128 g for Saudi-1, 79 to 130 g for Saudi-2, and 76 to 119 g for Saudi-3 along with moderate ratios of feed conversion ranging from 2.65 to 3.80, 2.45 to 3.90, 2.46 to 3.79 and 2.63 to 3.65, respectively. Pre-slaughter weight, hot carcass weight, and offal weight were in favour of Saudi-2 rabbits compared to the other groups. Both Saudi-1 and Saudi-2 rabbits were slightly higher than Saudi-3 in weights and percentages of head, fur, viscera and legs + tail. Lean and bone weights and percentages and meat to bone ratio in Saudi-2 carcasses were slightly higher than those recorded in the other groups, while moisture, DM, CP, EE and ash contents in the lean have shown little differences between groups. Rabbits of Saudi-1 were ranked the first in digestibility coefficients of OM, CP, NDF, ADF, HC, C and cell count compared to other groups.Rabbits of Saudi-2 fed diet containing 87.5% IFS recorded the heaviest body weights and gains since this class showed considerable deviations in body weights of 345, 341, 269, 307, 321, 345 and 347 g at 6, 7, 8, 9, 10, 11 and 12 weeks of age, respectively, in comparison with the lightest class. Both Saudi-2 and Saudi-3 rabbits fed the diet containing 87.5% IFS had favourable estimates of feed conversions ranging from 2.1 to 3.4, while rabbits of Saudi-1 fed the diet containing 87.5% IFS recorded the best digestibility coefficients.